Topical muscle relaxation compositions and methods

ABSTRACT

A method for reducing muscle contraction of a facial muscle or reducing the appearance of a fine line or wrinkle is disclosed. The method can include topical application of a composition to facial skin and/or to the fine line or wrinkle. The composition can include an effective amount of Rosmarinus officinalis leaf extract, an effective amount of Lavendula stoechas extract, and an effective amount of Acmella oleracea extract. Topical application of the composition to facial skin can reduce muscle contraction of the facial muscle. Topical application of the composition to the fine line or wrinkle can reduce the appearance of the fine line or wrinkle.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. patent applicationSer. No. 16/457,027 filed Jun. 28, 2019, which claims priority to U.S.Provisional Application No. 62/693,545 filed Jul. 3, 2018, thedisclosures of which are hereby incorporated by reference in theirentireties.

BACKGROUND OF THE INVENTION A. Field of the Invention

The present invention relates generally to a composition that can beused to relax facial muscles and/or reduce the appearance of fine linesor wrinkles. The composition can include a combination of plant-basedextracts such as Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract.

B. Background

Certain types of wrinkles develop over time on facial skin due torepetitive muscle contraction. These types of wrinkles are typicallyreferred to as rhytides, examples of which can include frown orglabellar lines, forehead lines, crow's feet, bunny or nasalis lines,dimpled chin, smile lift lines, lip lines, etc. Currently, a widevariety of commercially available treatment materials are available andare purportedly effective as facial muscle relaxants/muscle contractionsuppressants. These materials are said to be capable of reducing theappearance of such wrinkles. Non-limiting examples of such treatmentmaterials include injectable neuromodulators such as Botox®(onabotulinumtoxinA), Dysport® (abobotulinumtoxinA), or Xeomin®(incobotulinumtoxinA). Other examples include topically administeredtreatment materials such as chemical compounds (e.g.,gamma-aminobutyric-acid) and peptides (e.g., acetyl hexapeptide-3).

Various problems are associated with injectable neuromodulators, such asallergic reactions, rash, itching, injection site reactions (bruising,bleeding, pain, redness, swelling, or tenderness), muscle stiffness,fever, cough, sore throat, flu-like symptoms, neck or back pain, and thepotential to spread from the injection site to other parts of the body,resulting in serious risks including difficulty talking, swallowing orbreathing, muscle weakness, drooping eyelids, and blurred or doublevision. Similarly, chemical compounds have their own drawbacks such asskin irritation and potential allergic reactions. Aside from thephysiological problems associated with injectable neuromodulators andchemical compounds, these elective cosmetic procedures can becost-prohibitive for a large segment of the population who may benefit.

There have been a few attempts to produce natural-based ingredients thathave the ability to relax facial muscles. One such attempt is thedevelopment of Acmella oleracea extracts. While these extracts may offersome muscle relaxation effect, the effect may not be sufficient toeffectively relax facial muscles and reduce the appearance of fine linesor wrinkles.

Thus, previous attempts to improve the visual appearance of skin havebeen shown to have various drawbacks such as high costs, medical risks,skin irritation, prolonged recovery periods, or inefficient delivery ofthe promised skin benefits.

SUMMARY OF THE INVENTION

The inventors have identified a solution to at least some of theproblems associated with treating fine lines or wrinkles. The solutionresides in a combination of plant-based ingredients that is effective atreducing muscle contraction of facial muscles. In one particular aspect,the facial muscle contraction can be reduced by reducing contraction ofmyotubes via reducing an influx of calcium levels in the myotubes.Reducing the influx of calcium levels in the myotubes can reduce orinhibit an action potential in the myotubes, thus reducing or preventingmuscle contraction. This combination of ingredients also has the abilityto modulate biochemical pathways that can further help reduce theappearance of fine lines or wrinkles such as by stimulating collagenproduction in skin (e.g., keratinocytes or human dermal fibroblasts)and/or reducing enzymatic activity or production of enzymes associatedwith the degradation of extracellular matrix proteins (e.g., matrixmetalloproteinase (MMP) enzymes such as MMP-1, MMP3, and/or MMP-9).Still further, the combination of ingredients also has the ability toincrease collagen expression and laminin production in skin cells (e.g.,keratinocytes or human dermal fibroblasts). The combination of theplant-based ingredients include at least two of or all three ofRosmarinus officinalis leaf extract, Lavendula stoechas extract, and/orAcmella oleracea extract. This combination of plant-based ingredientscan be used to create topical skin compositions that reduce theappearance of and/or prevent the formation of fine lines or wrinkles,including deep facial line wrinkles or rhytides that can be linked torepetitive facial muscle contraction over prolonged periods of time(e.g., 1, 2, 4, 6, 8, 12, or more months or several years). Notably, thecombination of ingredients can be used to reduce the appearance ofand/or prevent the formation of both static and dynamic fine lines orwrinkles.

In one aspect of the present invention, there is disclosed a method forreducing muscle contraction of a facial muscle in a person. In anotheraspect, there is disclosed a method for reducing the appearance of afine line or wrinkle on a person's skin. The method can includetopically applying to skin a composition comprising: (a) an effectiveamount of Rosmarinus officinalis leaf extract; (b) an effective amountof Lavendula stoechas extract; and/or (c) an effective amount of Acmellaoleracea extract, wherein topical application of the composition tofacial skin reduces muscle contraction of the facial muscle or whereintopical application of the composition to skin reduces the appearance offine lines or wrinkles on a person's skin. Additionally, oralternatively, topical application can include rolling a micro-rolleracross the person's skin to increase penetration of the composition intothe skin. In certain aspects, the composition includes Rosmarinusofficinalis leaf extract and Lavendula stoechas extract, Rosmarinusofficinalis leaf extract and Acmella oleracea extract, Lavendulastoechas extract and Acmella oleracea extract, or all of Rosmarinusofficinalis leaf extract, Lavendula stoechas extract and Acmellaoleracea extract. In certain aspects, facial muscle(s) that is/arecontracted can be positioned directly under the skin where thecomposition is applied. In other aspects, facial muscle(s) that is/arecontracted can be in close proximity to the skin but not directlyunderneath the skin where the composition is applied (e.g., within 3centimeters, preferably with 2.5 cm, or more preferably within 2 cm, oreven more preferably within 1.5 or 1 cm of the area of the skin that thecomposition has been applied to). The facial muscle can be a glabellarcomplex muscle, an orbicularis oculi muscle, a depressor muscle, or afrontalis muscle, or any combination thereof. Alternatively, thecomposition can be applied to non-facial skin (e.g., hands, thighs,neck, buttocks, décolleté region, feet, etc.) and can lead to reducedmuscle contraction of muscles directly under the skin where thecomposition has been applied or in close proximity to the skin but notdirectly underneath the skin. In instances where the composition isapplied to a fine line or wrinkle, the fine line or wrinkle can be arhytide, and reduction in muscle contraction of the facial musclereduces the appearance of the rhytide. In other instances, the fine lineor wrinkle is not a rhytide.

The extracts used in the compositions of the present invention can eachindividually be obtained from water as an extracting solvent (e.g.,aqueous extract), alcohol as an extracting solvent (e.g., alcoholextract), or a polyol as the extracting solvent (e.g., polyol extract),or any combinations of such extracting solvents (e.g., aqueous-alcoholicextract, aqueous-polyol extract, alcohol-polyol extract oraqueous-alcoholic-polyol extract). Non-limiting examples of alcohols canbe methanol, ethanol, propanol, butanol, petnanol, hexanol, heptanol,octanol, etc. Non-limiting examples of polyols can be ethylene glycol,propylene glycol, glycerol, erythritol, xylitol, mannitol, volemitol,etc. Additionally, other extracting solvents can be used such asadditional hydrophilic solvents or lipophilic solvents (e.g., methane,ethane, butane, propane, hexane, heptane, octane, dimethyl sulfoxide(DMSO), N-methyl-2-pyrrolidone (NMP), carbon dioxide such as use ofcarbon dioxide (CO₂) in supercritical extraction techniques). CO₂supercritical extraction can include filling a column with ground driedplant material and pumping supercritical liquid carbon dioxide thoughthe column at very high pressure (200-400 Bar), and then collecting theresulting extract.

In a preferred instance, the Rosmarinus officinalis leaf extract can beobtained from the leaf of Rosmarinus officinalis. The leaf can besubjected to a eutectigenesis extraction process using a fluidextraction mixture comprising betaine or hydrated betaine, a hydrogenbond donor compound (e.g., polyols, organic acids, etc.), and water. Theeutectic extract can then be used in the compositions of the presentinvention. In some preferred instances, the hydrogen bond donor is anorganic acid, preferably lactic acid. Eutectigenesis utilizes eutecticsolvents which are mixtures of compounds having melting points lowerthan those of their constituents taken in isolation. In anotherpreferred instance, the Lavendula stoechas extract is from a combinationof the flower, leaf, and stem portions of Lavendula stoechas, and theextract can be prepared by subjecting these portions to a supercriticalextraction process using carbon dioxide (CO₂) as the solvent. Thesupercritical CO₂ extract of Lavendula stoechas flower, leaf, and stem(flower/leaf/stem) can then be used in the compositions of the presentinvention. The CO₂ supercritical extract of the Lavendula stoechasflower/leaf/stem can also be mixed with caprylic/capric triglyceridesand then used in the compositions of the present invention. In stillanother preferred instance, the Acmella oleracea extract is from thecombination of the flower, leaf, and stem portions of Acmella oleracea,and the extract can be prepared by subjecting these portions(flower/leaf/stem) to a hydro-alcoholic (preferably hydro-ethanolic)extraction process or a hydro-alcoholic-polyol extraction process. Thepolyol in preferred instances can be 1,3-propanediol. The alcohol can beremoved from the resulting extract. The Acmella oleracea extract can bethen be used in the compositions of the present invention. Thus, in apreferred instance, the Rosmarinus officinalis leaf eutectic extract,the Lavendula stoechas flower/leaf/stem supercritical CO₂ extract, andthe Acmella oleracea flower/leaf/stem hydro-alcohol-polyol extract canbe used in the compositions and methods of the present invention.

An effective amount of Rosmarinus officinalis leaf extract in thecomposition can be 0.00001 to 25% w/w or any range therein (e.g.,0.00001 to 10% w/w, 0.00001 to 5% w/w, 0.00001 to 2% w/w, 0.00001 to 1%w/w, 0.00001 to 0.5% w/w, or 0.001 to 1% w/w, 0.01 to 1% w/w, 0.1 to 1%w/w. or 0.001 to 2% w/w, 0.01 to 2% w/w, or 0.1 to 2% w/w). An effectiveamount of Lavendula stoechas extract in the composition can be 0.00001to 25 w/w or any range therein (e.g., 0.00001 to 10% w/w, 0.00001 to 5%w/w, 0.00001 to 2% w/w, 0.00001 to 1% w/w, 0.00001 to 0.5% w/w, or 0.001to 1% w/w, 0.01 to 1% w/w, 0.1 to 1% w/w. or 0.001 to 2% w/w, 0.01 to 2%w/w, or 0.1 to 2% w/w). An effective amount of Acmella oleracea extractin the composition can be 0.00001 to 25% w/w or any range therein (e.g.,0.00001 to 10% w/w, 0.00001 to 5% w/w, 0.00001 to 2% w/w, 0.00001 to 1%w/w, 0.00001 to 0.5% w/w, or 0.001 to 1% w/w, 0.01 to 1% w/w, 0.1 to 1%w/w. or 0.001 to 2% w/w, 0.01 to 2% w/w, or 0.1 to 2% w/w).

The compositions of the present invention can reduce contraction ofmyotubes. This can be done by reducing the influx of calcium into themyotubes, which can reduce or prevent an action potential from occurringin the myotubes. By reducing or preventing action potential development,contraction of the myotubes can be reduced or prevented. Notably, eachof Rosmarinus officinalis leaf or an extract thereof, Lavendula stoechasor an extract thereof, and/or Acmella oleracea or an extract thereof hasbeen shown by the inventors to reduce or prevent an influx of calciuminto myotubes and to reduce or prevent muscle contraction. Thecompositions of the present invention can also stimulate collagenproduction in skin cells (e.g., keratinocytes and/or human dermalfibroblasts). In particular, it was discovered in the context of thepresent invention that Acmella oleracea or an extract thereof canincrease collagen production in skin cells. The compositions of thepresent invention can also stimulate laminin production in skin cells(e.g., keratinocytes and/or human dermal fibroblasts). In particular, itwas discovered in the context of the present invention that Acmellaoleracea or an extract thereof can increase laminin production in skincells. The compositions of the present invention can also reduce orinhibit matrix metalloproteinases (MMP) 1, 3, and 9 activity orproduction in skin cells (e.g., keratinocytes and/or human dermalfibroblasts). In particular, it was discovered in the context of thepresent invention that the Rosmarinus officinalis leaf or an extractthereof can reduce or inhibit each of MMP-1, MMP-3, and MMP-9 activityor production in skin cells. The skin can be facial skin such as skinlocated on a person's forehead, cheeks, chin, orbital area region (e.g.,under the eyes, adjacent to the eyes such as where crow's feet form,eyelid, etc.). In certain preferred aspects, the facial skin is foreheadskin or orbital area region skin, especially where crow's feet develop.Still further, the compositions of the present invention can be used toincrease skin firmness, which can aid in reducing the appearance ofloos, sagging, and/or flaccid skin.

In addition to Rosmarinus officinalis leaf or an extract thereof,Lavendula stoechas or an extract thereof, and/or Acmella oleracea or anextract thereof, the topical skin compositions of the present inventioncan further comprise one or more ingredients described herein. Forexample, the composition can comprise one or more additional ingredientsselected from one or more cosmetic ingredients or pharmaceuticalingredients. In some instances, the compositions can include one or moreconditioning agents, moisturizing agents, pH adjusters, structuringagents, silicone containing compounds, inorganic salts, and/orpreservatives. In some aspects, the topical composition further includeswater, preferably at least 50, 60, 70, 80, or 90, or more wt. % water,more preferably between 50 to 90 wt. % water. In some aspects, thetopical compositions of the present invention may exclude one or moreadditional ingredients selected from one or more cosmetic ingredients orpharmaceutical ingredients. In still some particular aspects, thecompositions of the present invention may exclude one or more ofRosmarinus officinalis leaf extract, Lavendula stoechas extract, andAcmella oleracea extract. In some aspects, the topical compositionexcludes water. In some aspects, the topical composition herein may beanhydrous or substantially anhydrous.

In particular aspects, the compositions of the present invention areformulated as a topical skin composition. The composition can have adermatologically acceptable vehicle or carrier for the compounds andextracts. The composition can further include a moisturizing agent or ahumectant, a surfactant, a silicone containing compounds, a UV agent, anoil, and/or other ingredients identified in this specification or thoseknown in the art. The composition can be a mask, lotion, cream, gel,serum, emulsion (e.g., oil-in-water, water-in-oil, silicone-in-water,water-in-silicone, water-in-oil-in-water, oil-in-water-in-oil,oil-in-water-in-silicone, etc.), solutions (e.g., aqueous orhydro-alcoholic solutions), anhydrous bases (e.g., lipstick or apowder), ointments, milk, paste, aerosol, solid forms, eye jellies, gelserums, gel emulsions, etc. The composition can be formulated fortopical skin application at least 1, 2, 3, 4, 5, 6, 7, or more times aday during use.

In other aspects of the present invention, compositions can be storagestable or color stable, or both. It is also contemplated that theviscosity of the composition can be selected to achieve a desiredresult, e.g., depending on the type of composition desired, theviscosity of such composition can be from about 1 cps to well over 1million cps or any range or integer derivable therein (e.g., 2 cps, 3,4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300,400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000, 6000, 7000,8000, 9000, 10000, 20000, 30000, 40000, 50000, 60000, 70000, 80000,90000, 100000, 200000, 300000, 400000, 500000, 600000, 700000, 800000,900000, 1000000, 2000000, 3000000, 4000000, 5000000, 10000000, cps,etc., as measured on a Brookfield Viscometer using a TC spindle at 2.5rpm at 25° C.).

The compositions in non-limiting aspects can have a pH of about 6 toabout 9. In other aspects, the pH can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, or 14. The compositions can include a triglyceride.Non-limiting examples include small, medium, and large chaintriglycerides. In certain aspects, the triglyceride is a medium chaintriglyceride (e.g., caprylic capric triglyceride). The compositions canalso include preservatives. Non-limiting examples of preservativesinclude sodium benzoate, iodopropynyl butylcarbamate, methylparaben,propylparaben, or a mixture of methylparaben and propylparaben. In someembodiments, the composition is paraben-free.

Compositions of the present invention can have UVA and UVB absorptionproperties. The compositions can have a sun protection factor (SPF) of2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45,50, 55, 60, or more, or any integer or derivative therein. Thecompositions can be sunscreen lotions, sprays, or creams.

The compositions of the present invention can also include any one of,any combination of, or all of the following additional ingredients:water, a chelating agent, a moisturizing agent, a preservative, athickening agent, a silicone containing compound, an essential oil, astructuring agent, a vitamin, a pharmaceutical ingredient, anantioxidant, or any combination of such ingredients or mixtures of suchingredients. In certain aspects, the composition can include at leasttwo, three, four, five, six, seven, eight, nine, ten, or all of theseadditional ingredients identified in the previous sentence. Non-limitingexamples of these additional ingredients are identified throughout thisspecification and are incorporated into this section by reference. Theamounts of such ingredients can range from 0.0001% to 99.9% by weight orvolume of the composition, or any integer or range in between asdisclosed in other sections of this specification, which areincorporated into this paragraph by reference.

Kits that include the compositions of the present invention are alsocontemplated. In certain embodiments, the composition is comprised in acontainer. The container can be a bottle, dispenser, or package. Thecontainer can dispense a pre-determined amount of the composition. Incertain aspects, the compositions is dispensed in a spray, mist, dollop,or liquid. The container can include indicia on its surface. The indiciacan be a word, an abbreviation, a picture, or a symbol.

It is also contemplated that the compositions disclosed throughout thisspecification can be used as a leave-on or rinse-off composition. By wayof example, a leave-on composition can be one that is topically appliedto skin and remains on the skin for a period of time (e.g., at least 5,6, 7, 8, 9, 10, 20, or 30 minutes, or at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours,or overnight or throughout the day). Alternatively, a rinse-offcomposition can be a product that is intended to be applied to the skinand then removed or rinsed from the skin (e.g., with water) within aperiod of time such as less than 5, 4, 3, 2, or 1 minute. An example ofa rinse off composition can be a skin cleanser, shampoo, conditioner, orsoap. An example of a leave-on composition can be a skin moisturizer,sunscreen, mask, overnight cream, or a day cream.

It is contemplated that any embodiment discussed in this specificationcan be implemented with respect to any method or composition of theinvention, and vice versa. Furthermore, compositions of the inventioncan be used to achieve methods of the invention.

In one embodiment, compositions of the present invention can bepharmaceutically or cosmetically elegant or can have pleasant tactileproperties. “Pharmaceutically elegant,” “cosmetically elegant,” and/or“pleasant tactile properties” describes a composition that hasparticular tactile properties which feel pleasant on the skin (e.g.,compositions that are not too watery or greasy, compositions that have asilky texture, compositions that are non-tacky or sticky, etc.).Pharmaceutically or cosmetically elegant can also relate to thecreaminess or lubricity properties of the composition or to the moistureretaining properties of the composition.

Also contemplated is a product comprising a composition of the presentinvention. In non-limiting aspects, the product can be a cosmeticproduct. The cosmetic product can be those described in other sectionsof this specification or those known to a person of skill in the art.Non-limiting examples of products include a moisturizer, a cream, alotion, a skin softener, a gel, a wash, a foundation, a night cream, alipstick, a cleanser, a cleansing balm, a toner, a sunscreen, a mask, ananti-aging product, a deodorant, an antiperspirant, a perfume, acologne, etc.

Also disclosed are the following Embodiments 1 to 40 of the presentinvention. Embodiment 1 is a method for reducing muscle contraction of afacial muscle in a person, the method comprising topically applying tofacial skin a composition comprising: (a) an effective amount ofRosmarinus officinalis leaf extract; (b) an effective amount ofLavendula stoechas extract; and (c) an effective amount of Acmellaoleracea extract, wherein topical application of the composition to thefacial skin reduces muscle contraction of the facial muscle. Embodiment2 is the method of claim 1, wherein the composition is applied to a fineline or wrinkle, and wherein the appearance of the fine line or wrinkleis reduced after topical application. Embodiment 3 is the method ofclaim 2, wherein the fine line or wrinkle is a rhytide, and wherein thereduction in muscle contraction of the facial muscle reduces theappearance of the rhytide. Embodiment 4 is the method of any one ofEmbodiments 1 to 3, wherein the facial muscle is a glabellar complexmuscle, an orbicularis oculi muscle, a depressor muscle, or a frontalismuscle, or any combination thereof. Embodiment 5 is the method of anyone of Embodiments 1 to 4, wherein each of the extracts is individuallyan aqueous extract, an alcohol extract, a polyol extract, or acombination thereof. Embodiment 6 is the method of any one ofEmbodiments 1 to 5, wherein: the Rosmarinus officinalis leaf extract isobtained with a fluid extraction solvent mixture comprising betaine,lactic acid, and water; the Lavendula stoechas extract is a Lavendulastoechas flower/leaf/stem extract obtained with a supercritical carbondioxide (CO₂) extraction solvent; and the Acmella oleracea extract isobtained with a fluid extraction solvent mixture comprising water,ethanol, and 1,3 propanediol. Embodiment 7 is the method of any ofEmbodiments 1 to 6, wherein the composition comprises: 0.00001 to 10%w/w of Rosmarinus officinalis leaf extract; 0.00001 to 10% w/w ofLavendula stoechas extract; and, 0.00001 to 10% w/w of Acmella oleraceaextract. Embodiment 8 is the method of any of Embodiments 1 to 7,wherein: the Rosmarinus officinalis leaf extract reduces an influx ofcalcium in the facial muscle and reduces the occurrence of an actionpotential in the facial muscle; the Lavendula stoechas extract reducesan influx of calcium in the facial muscle and reduces the occurrence ofan action potential in the facial muscle; and the Acmella oleraceaextract reduces an influx of calcium in the facial muscle and reducesthe occurrence of an action potential in the facial muscle. Embodiment 9is the method of any one of Embodiments 1 to 8, wherein the Acmellaoleracea extract stimulates collagen production and laminin productionin the skin. Embodiment 10 is the method of any one of Embodiments 1 to9, wherein the Rosmarinus officinalis leaf extract reduces matrixmetalloproteinases 1, 3, or 9 production in the skin. Embodiment 11 isthe method of any one of Embodiments 1 to 10, wherein the facial skin isforehead skin, cheek skin, chin skin, or orbital area skin. Embodiment12 is the method of Embodiment 11, wherein the skin is forehead skin.Embodiment 13 is the method of Embodiment 11, wherein the skin isorbital area skin. Embodiment 14 is the method of any of Embodiments 1to 13, wherein the composition is an emulsion. Embodiment 15 is themethod of Embodiment 14, wherein the emulsion is an oil-in-wateremulsion. Embodiment 16 is the method of any one of Embodiments 1 to 13,wherein the composition is a gel. Embodiment 17 is a method of reducingthe appearance of a fine line or wrinkle on a person's skin, the methodcomprising topically applying to the fine line or wrinkle a compositioncomprising: (a) an effective amount of Rosmarinus officinalis leafextract; (b) an effective amount of Lavendula stoechas extract; and (c)an effective amount of Acmella oleracea extract, wherein topicalapplication reduces the appearance of the fine line or wrinkle.Embodiment 18 is the method of Embodiment 17, wherein the compositionreduces muscle contraction of a muscle that causes the appearance of thefine line or wrinkle when the muscle is contracted. Embodiment 19 is themethod of any one of Embodiments 17 to 18, wherein the muscle is aglabellar complex muscle, an orbicularis oculi muscle, a depressormuscle, or a frontalis muscle, or any combination thereof. Embodiment 20is the method of any one of Embodiments 17 to 19, wherein the fine lineor wrinkle is a rhytide. Embodiment 21 is the method of any one ofEmbodiments 17 to 20, wherein each of the extracts is individually anaqueous extract, an alcohol extract, a polyol extract, or a combinationthereof. Embodiment 22 is the method of any one of Embodiments 17 to 21,wherein: the Rosmarinus officinalis leaf extract is obtained with afluid extraction solvent mixture comprising betaine, lactic acid, andwater; the Lavendula stoechas extract is a Lavendula stoechasflower/leaf/stem extract obtained with a supercritical carbon dioxide(CO₂) extraction solvent; and the Acmella oleracea extract is obtainedwith a fluid extraction solvent mixture comprising water, ethanol, and1,3 propanediol. Embodiment 23 is the method of any of Embodiments 17 to22, wherein the composition comprises: 0.00001 to 10% w/w of Rosmarinusofficinalis leaf extract; 0.00001 to 10% w/w of Lavendula stoechasextract; and, 0.00001 to 10% w/w of Acmella oleracea extract. Embodiment24 is the method of any of Embodiments 17 to 23, wherein: the Rosmarinusofficinalis leaf extract reduces an influx of calcium in the facialmuscle and reduces the occurrence of an action potential in the facialmuscle; the Lavendula stoechas extract reduces an influx of calcium inthe facial muscle and reduces the occurrence of an action potential inthe facial muscle; and the Acmella oleracea extract reduces an influx ofcalcium in the facial muscle and reduces the occurrence of an actionpotential in the facial muscle. Embodiment 25 is the method of any oneof Embodiments 17 to 24, wherein the Acmella oleracea extract stimulatescollagen production and laminin production in the skin. Embodiment 26 isthe method of any one of Embodiments 17 to 25, wherein the Rosmarinusofficinalis leaf extract reduces matrix metalloproteinases 1, 3, or 9production in the skin. Embodiment 27 is the method of any one ofEmbodiments 17 to 26, wherein the person's skin is facial skin,preferably forehead skin, cheek skin, chin skin, or orbital area skin.Embodiment 28 is the method of Embodiment 27, wherein the skin isforehead skin. Embodiment 29 is the method of Embodiment 27, wherein theskin is orbital area skin. Embodiment 30 is the method of any ofEmbodiments 17 to 29, wherein the composition is an emulsion. Embodiment31 is the method of Embodiment 30, wherein the emulsion is anoil-in-water emulsion. Embodiment 32 is the method of any one ofEmbodiments 17 to 31, wherein the composition is a gel. Embodiment 33 isa topical skin composition comprising: (a) an effective amount ofRosmarinus officinalis leaf extract; (b) an effective amount ofLavendula stoechas extract; and (c) an effective amount of Acmellaoleracea extract, wherein the topical skin composition is capable ofreducing the appearance of a fine line or wrinkle in a person's skinand/or is capable of reducing muscle contraction of a facial muscle in aperson. Embodiment 34 is the composition of Embodiment 33, wherein eachof the extracts is individually an aqueous extract, an alcohol extract,a polyol extract, or a combination thereof. Embodiment 35 is thecomposition of any one of Embodiments 33 to 34, wherein: the Rosmarinusofficinalis leaf extract is obtained with a fluid extraction solventmixture comprising betaine, lactic acid, and water; the Lavendulastoechas extract is a Lavendula stoechas flower/leaf/stem extractobtained with a supercritical carbon dioxide (CO₂) extraction solvent;and the Acmella oleracea extract is obtained with a fluid extractionsolvent mixture comprising water, ethanol, and 1,3 propanediol.Embodiment 36 is the composition of any one of Embodiment 33 to 35,wherein the composition comprises: 0.00001 to 10% w/w of Rosmarinusofficinalis leaf extract; 0.00001 to 10% w/w of Lavendula stoechasextract; and, 0.00001 to 10% w/w of Acmella oleracea extract. Embodiment37 is the composition of any one of Embodiments 33 to 36, wherein: theRosmarinus officinalis leaf extract is capable of reducing an influx ofcalcium in the facial muscle and reduces the occurrence of an actionpotential in the facial muscle; the Lavendula stoechas extract iscapable of reducing an influx of calcium in the facial muscle andreduces the occurrence of an action potential in the facial muscle; andthe Acmella oleracea extract is capable of reducing an influx of calciumin the facial muscle and reduces the occurrence of an action potentialin the facial muscle. Embodiment 38 is the composition of any one ofEmbodiments 33 to 37, wherein the Acmella oleracea extract is capable ofstimulating collagen production and laminin production in the skin.Embodiment 39 is the composition of any one of Embodiments 33 to 38,wherein the Rosmarinus officinalis leaf extract is capable of reducingmatrix metalloproteinases 1, 3, or 9 production in the skin. Embodiment40 is the composition of any one of Embodiments 33 to 39, wherein thecomposition is an emulsion, preferably an oil-in-water emulsion, or agel.

“Topical application” means to apply or spread a composition onto thesurface of lips or keratinous tissue. “Topical skin composition”includes compositions suitable for topical application on skin and/orkeratinous tissue. Such compositions are typicallydermatologically-acceptable in that they do not have undue toxicity,incompatibility, instability, allergic response, and the like, whenapplied to skin and/or keratinous tissue. Topical skin care compositionsof the present invention can have a selected viscosity to avoidsignificant dripping or pooling after application to skin and/orkeratinous tissue.

“Keratinous tissue” includes keratin-containing layers disposed as theoutermost protective covering of mammals and includes, but is notlimited to, lips, skin, hair, and nails.

The term “about” or “approximately” are defined as being close to asunderstood by one of ordinary skill in the art. In one non-limitingembodiment the terms are defined to be within 10%, preferably within 5%,more preferably within 1%, and most preferably within 0.5%.

The term “substantially” and its variations refer to ranges within 10%,within 5%, within 1%, or within 0.5%.

The terms “inhibiting” or “reducing” or any variation of these termsincludes any measurable decrease or complete inhibition to achieve adesired result. The terms “promote” or “increase” or any variation ofthese terms includes any measurable increase or production of a proteinor molecule (e.g., matrix proteins such as fibronectin, laminin,collagen, or elastin or molecules such as hyaluronic acid) to achieve adesired result.

The term “effective,” as that term is used in the specification and/orclaims, means adequate to accomplish a desired, expected, or intendedresult.

The use of the word “a” or “an” when used in conjunction with the terms“comprising,” “including,” “having,” or “containing,” or any variationsof these terms, in the claims and/or the specification may mean “one,”but it is also consistent with the meaning of “one or more,” “at leastone,” and “one or more than one.”

The terms “wt. %,” “vol. %,” or “mol. %” refers to a weight, volume, ormolar percentage of a component, respectively, based on the totalweight, the total volume, or the total moles of material that includesthe component. In a non-limiting example, 10 grams of component in 100grams of the material is 10 wt. % of component. The term “% w/w” has thesame meaning as wt. %.

As used in this specification and claim(s), the words “comprising” (andany form of comprising, such as “comprise” and “comprises”), “having”(and any form of having, such as “have” and “has”), “including” (and anyform of including, such as “includes” and “include”) or “containing”(and any form of containing, such as “contains” and “contain”) areinclusive or open-ended and do not exclude additional, unrecitedelements or method steps.

The compositions and methods for their use can “comprise,” “consistessentially of,” or “consist of” any of the ingredients or stepsdisclosed throughout the specification. With respect to the phrase“consisting essentially of,” a basic and novel property of thecompositions and methods of the present invention is the ability toreduce the appearance of fine lines or wrinkles and/or reduce musclecontraction, preferably facial muscle contraction.

Other objects, features, and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the examples,while indicating specific embodiments of the invention, are given by wayof illustration only. Additionally, it is contemplated that changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed description ofthe specification embodiments presented herein.

FIG. 1 provides data illustrating the mean % of improvement frombaseline (0 week) in the EMG response area (mV.s) for muscle contractionin the glabella region for a group of people using a composition of thepresent invention having a combination of Rosmarinus officinalis leafextract, Lavendula stoechas extract, and Acmella oleracea extract. Themean % of improvement illustrates reduced muscle contraction in theglabella region over time (2 weeks, 4 weeks, and 8 weeks). “*” indicatesp<0.05 statistically significant improvement when compared to baseline(0 weeks).

FIG. 2 provides data illustrating the % of people showing improvementfrom baseline (0 week) in the EMG response area (mV.s) for musclecontraction in the glabella region for a group of people using acomposition of the present invention having a combination of Rosmarinusofficinalis leaf extract, Lavendula stoechas extract, and Acmellaoleracea extract. The % of people showing improvement illustratesreduced muscle contraction in the glabella region over time (2 weeks, 4weeks, and 8 weeks).

FIG. 3 provides data illustrating the mean % of improvement frombaseline (0 week) for reduced appearance of fine lines and wrinkles fora group of people using a composition of the present invention having acombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract. The mean % of improvementillustrates reduced appearance of fine lines and wrinkles in the treatedarea over time (2 weeks, 4 weeks, and 8 weeks). “*” indicates p<0.05statistically significant improvement when compared to baseline (0weeks).

FIG. 4 provides data illustrating the % of people showing improvementfrom baseline (0 week) in the reduction of fine lines and wrinkles for agroup of people using a composition of the present invention having acombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract. The % of people showingimprovement illustrates reduced appearance of fine lines and wrinkles inthe treated area over time (2 weeks, 4 weeks, and 8 weeks).

FIG. 5 provides data illustrating the mean % of improvement frombaseline (0 week) in the EMG response area (mV.$) for muscle contractionin the glabella region for a group of people using a composition of thepresent invention having a combination of Rosmarinus officinalis leafextract, Lavendula stoechas extract, and Acmella oleracea extract, andapplying the composition with a micro-roller versus applying thecomposition without a micro-roller. The mean % of improvementillustrates a faster and stronger reduction in facial muscle contractionat 2, 4, and 8 weeks compared to baseline when using a micro-roller withapplication of the composition. “*” indicates p<0.05 statisticallysignificant improvement when compared to baseline (0 weeks).

FIG. 6 provides data illustrating the % of people showing improvementfrom baseline (0 week) in the EMG response area (mV.s) for musclecontraction in the glabella region for a group of people using acomposition of the present invention having a combination of Rosmarinusofficinalis leaf extract, Lavendula stoechas extract, and Acmellaoleracea extract. The % of people showing improvement illustrates afaster and stronger reduction in facial muscle contraction at 2, 4, and8 weeks in the glabella region when using a micro-roller withapplication of the composition.

FIG. 7 provides data illustrating the mean % of improvement frombaseline (0 week) for reduced appearance of fine lines and wrinkles fora group of people using a composition of the present invention having acombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract. The mean % of improvementillustrates reduced appearance of fine lines and wrinkles in the treatedarea over time (2 weeks, 4 weeks, and 8 weeks) when using a micro-rollerwith application of the composition and without using a micro-rollerwith application of the composition. “*” indicates p<0.05 statisticallysignificant improvement when compared to baseline (0 weeks).

FIG. 8 provides data illustrating the % of people showing improvementfrom baseline (0 week) in the reduction of fine lines and wrinkles for agroup of people using a composition of the present invention having acombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract. The % of people showingimprovement illustrates reduced appearance of fine lines and wrinkles inthe treated area over time (2 weeks, 4 weeks, and 8 weeks) when using amicro-roller with application of the composition and without using amicro-roller with application of the composition.

FIG. 9 provides data illustrating the mean % of improvement frombaseline (0 week) for skin texture/roughness for a group of people usinga composition of the present invention having a combination ofRosmarinus officinalis leaf extract, Lavendula stoechas extract, andAcmella oleracea extract. The mean % of improvement illustrates improvedskin texture/roughness in the treated area over time (2 weeks, 4 weeks,and 8 weeks) when using a micro-roller with application of thecomposition and without using a micro-roller with application of thecomposition. “*” indicates p<0.05 statistically significant improvementwhen compared to baseline (0 weeks).

FIG. 10 provides data illustrating the % of people showing improvementfrom baseline (0 week) in skin texture/roughness for a group of peopleusing a composition of the present invention having a combination ofRosmarinus officinalis leaf extract, Lavendula stoechas extract, andAcmella oleracea extract. The % of people showing improvementillustrates improved skin texture/roughness in the treated area overtime (2 weeks, 4 weeks, and 8 weeks) when using a micro-roller withapplication of the composition and without using a micro-roller withapplication of the composition.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

A discovery has been made that provides a solution to at least some ofthe problems associated with current methods that are used to treat finelines or wrinkles in skin, including both static and dynamic fine linesor wrinkles. Fine lines are typically characterized as smaller andshallower (e.g., generally less than two millimeters in depth) whencompared to wrinkles, which have deeper lines (e.g., generally two ormore millimeters in depth). Static fine lines or wrinkles are typicallyassociated with fine lines or wrinkles that are present on skin that isrelaxed/not-moving. By comparison, dynamic fine lines or wrinkles appearon skin with skin movement associated with muscle contraction. Thesolution is premised on using a combination of at least two of, andpreferably, all three of, Rosmarinus officinalis leaf extract, Lavendulastoechas extract, and Acmella oleracea extract in topical skincompositions to reduce muscle contraction, preferably reduce musclecontraction in facial muscles. It is believed that the combination ofthese ingredients can help reduce the appearance of static and/ordynamic fine lines or wrinkles on at least three levels: (1) reducingmuscle contraction in instances where muscle contraction can form thepresence of fine lines or wrinkles (e.g., dynamic fine lines orwrinkles); (2) increasing collagen production in skin (e.g., in skincells such as keratinocytes and/or fibroblasts), which can allow formore full and firm skin, which can lead to reduced appearance of staticand/or dynamic fine lines or wrinkles; (3) decreasing matrixmetalloproteinase (MMP) production or activity (the terms “production”and “activity” can be used interchangeably throughout thisspecification) in skin such as MMP-1, MMP-3, and/or MMP-9 production oractivity to further help reduce the appearance of static and/or dynamicfine lines or wrinkles—MMP enzymes can breakdown or degradeextracellular matrix components such as collagen, fibronectin, laminin;proteoglycans, and vitronectin, which can lead to the formation of finelines or wrinkles; and (4) increase laminin production in skin (e.g.,skin cells such as keratinocytes and/or fibroblasts)—laminin is astructural glycoproteins located in the DEJ. Considered a glue thatholds the cells together, laminin is secreted by dermal fibroblasts tohelp facilitate intra- and inter-cellular adhesion of the epidermalcells to the DEJ.

These and other non-limiting aspects of the present invention aredescribed in the following sections.

A. Active Ingredients

The present invention is premised on a determination that a combinationof active ingredients—Rosmarinus officinalis leaf extract, Lavendulastoechas extract, and/or Acmella oleracea extract—can be used to reducemuscle contraction of a facial muscle, reduce and/or prevent facial finelines or wrinkles, reduce dynamic facial wrinkles and forehead wrinklesthat are caused due to repetitive facial muscle contraction (e.g.,rhytides), increase collagen production, reduce a matrixmetalloproteinase, including one or more of matrix metalloproteinase 1,matrix metalloproteinase 3, and matrix metalloproteinase 9, firm skin,and/or reduce the appearance of loose, sagging, and flaccid skin.

Rosmarinus officinalis leaf extract is an extract from the leaf ofRosmarinus officinalis. Rosmarinus officinalis is native to theMediterranean region, and is a woody, perennial herb with fragrant,evergreen, needle-like leaves and white, pink, purple, or blue flowers.It is a shrub that can reach up to 1.5 meters in height with leaves thatare about 2 to 4 cm long with green (top surface) and white (bottomsurface) coloring. In a preferred instance, the Rosmarinus officinalisleaf extract can be obtained from the leaf of Rosmarinus officinalis.The leaf can be subjected to a eutectigenesis extraction process using afluid extraction mixture comprising betaine or hydrated betaine, ahydrogen bond donor compound (e.g., polyols, organic acids, etc.), andwater. In particular, the leaf portion can be crushed or macerated andthen subjected to the aforementioned eutectic fluid extraction mixtureto obtain a eutectic extract. The eutectic extract can then be used inthe compositions of the present invention. In some preferred instances,the hydrogen bond donor is an organic acid, preferably lactic acid.Eutectigenesis utilizes eutectic solvents which are mixtures ofcompounds having melting points lower than those of their constituentstaken in isolation. In some instances, Rosmarinus officinalis iscommercially available. In some instances, Rosmarinus officinalis can besupplied by Naturex (France) under the trade name ROSEMARY EUTECTYSBLA™. It was discovered in the context of the present invention thatRosmarinus officinalis leaf extract can reduce or inhibit contraction ofmyotubes by reducing or inhibiting an influx of calcium in the myotubesand reducing or preventing the occurrence of an action potential. It wasalso discovered that Rosmarinus officinalis leaf extract can inhibit orreduce MMP-1, MMP-3, and MMP-3 production or activity in skin cells(e.g., keratinocytes or human dermal fibroblasts).

Lavendula stoechas extract is an extract from the plant Lavendulastoechas. Lavendula stoechas is native to the Mediterranean region, andis an evergreen shrub that is about 30 to 100 cm tall with greyish andtomentose leaves that are about 1 to 4 cm long. In a preferred instance,the Lavendula stoechas extract is from a combination of the flower,leaf, and stem portions of Lavendula stoechas. These portions can becombined and then crushed or macerated or crushed or macerated and thencombined. The resulting crushed or macerated flower/leaf/stem materialcan then subjected to a supercritical extraction process using carbondioxide (CO₂) as the solvent. The supercritical CO₂ extract of Lavendulastoechas flower/leaf/stem can then be used in the compositions of thepresent invention. The CO₂ supercritical extract of the Lavendulastoechas flower/leaf/stem can also be mixed with caprylic/caprictriglycerides and then used in the compositions of the presentinvention. In some instances, Lavendula stoechas extract is commerciallyavailable. In some instances, Lavendula stoechas can be supplied byBarnet Products LLC (Englewood Cliffs, N.J. (USA)) under the trade nameSTOCHEY'S. It was discovered in the context of the present inventionthat Lavendula stoechas extract can reduce or inhibit contraction ofmyotubes by reducing or inhibiting an influx of calcium in the myotubesand reducing or preventing the occurrence of an action potential.

Acmella oleracea extract is an extract from the Acmella oleracea plant.Acmella oleracea can be found in South America, Madagascar, and theMascarene Islands. In a preferred instance, the Acmella oleracea extractis from the combination of the flower, leaf, and stem portions ofAcmella oleracea. These portions can be combined and then crushed ormacerated or crushed or macerated and then combined. The resultingcrushed or macerated flower/leaf/stem material can then be subjected toa hydro-alcoholic (preferably hydro-ethanolic) extraction process or ahydro-alcoholic-polyol extraction process. The polyol in preferredinstances can be 1,3-propanediol. The alcohol can be removed from theresulting extract. The Acmella oleracea extract can be then be used inthe compositions of the present invention. In a preferred instance, theAcmella oleracea flower/leaf/stem extract can be obtained from ahydro-ethanol-1,3-propanediol solvent mixture. The resulting extract canthen be used in the compositions of the present invention or can befurther processed to remove the ethanol and can then be used in thecompositions of the present invention. Alternatively, the extractingsolvent can be a combination of water and polyol, preferably1,3-propanediol, without an alcohol. The amounts of water, alcohol,and/or polyol present in the reaction mixture can be modified asdesired. In some instances, Acmella oleracea extract is commerciallyavailable. In some instances, Acmella oleracea extract can be suppliedby Gattefosse (France) under the trade name GATULINE® EXPRESSION AF. Itwas discovered in the context of the present invention that Acmellaoleracea extract can reduce or inhibit contraction of myotubes byreducing or inhibiting an influx of calcium in the myotubes and reducingor preventing the occurrence of an action potential. It was alsodiscovered that Acmella oleracea extract can increase both collagenproduction and laminin production in skin cells (e.g., keratinocytes orhuman dermal fibroblasts).

The extracts described herein can be extracts made through extractionmethods known in the art and combinations thereof. Non-limiting examplesof extraction methods include the use of liquid-liquid extraction, solidphase extraction, aqueous extraction, ethyl acetate, alcohol, acetone,oil, supercritical carbon dioxide, heat, pressure, pressure dropextraction, ultrasonic extraction, etc. Extracts can be a liquid, solid,dried liquid, re-suspended solid, etc. In certain instances, each of theextracts can be prepared by: (i) obtaining the desired plant part (e.g.,leaf, stem, bark, flower, seed, etc.) or whole plant; (ii) crushing ormacerating the plant part or whole plant; (iii) optionally drying thecrushed or macerated plant part or whole plant; (iv) subjecting thecrushed or macerated plant or plant part to an extraction solvent to asufficient period of time (e.g., 1, 5, 10, 30, or 45 minutes or more, or1, 6, 12, or 18 hours or more, or 2, 3, 4, 5, 6 days, or more) underroom temperature (e.g., 20 to 30 ° C.) or heated (e.g., greater than 30° C. or more, preferably 30 ° C. to less than the boiling point of thesolvent); (v) collecting the solution comprising the extracting solventand the extracted plant material (e.g., liquid extract); and (vi)optionally removing the extracting solvent to obtain a dried plantextract; and (vii) optionally reconstituting the dried plant extract ina liquid carrier (e.g., water, alcohol, diol, etc.). It is alsocontemplated in the context of the present invention that plant materialfrom Rosmarinus officinalis leaf, Lavendula stoechas, and/or the Acmellaoleracea can be directly used without subjecting the plant material toan extraction technique.

Non-limiting examples of facial muscles in which the compositions of thepresent invention can attenuate contraction of include facial muscles ofthe glabellar complex, orbicularis muscles, frontalis muscles, ordepressor anguli oris muscles, or combinations thereof. The muscles ofthe glabellar complex can be responsible for formation of frown linesthat include the corrugator supercilii, depressor supercilii, procerus,and orbicularis oculi para frontalis muscles. The corrugator superciliilies below the frontalis muscle and functions to draw the brow mediallyand downward, whereas the smaller depressor supercilii can be locatedlower than the corrugator muscle and functions to draw the medial browdownward. The procerus can be located between the eyebrows and alsoworks to depress the glabellar medial brow region. The depressorfunction of the glabellar complex is opposed by the frontalis muscle.This muscle is merged with the superior portions of the glabellarcomplex, from which it extends upwards underneath the forehead. Thefrontalis muscle is a muscle of the forehead and is associated withraising of the brow. The orbicularis muscle is a muscle that closes theeyelids. The depressor anguli oris muscle is a facial muscle associatedwith frowning. In a preferred aspect, contractile activity is measured.

B. Amounts of Ingredients

It is contemplated that the compositions of the present invention caninclude any amount of the ingredients discussed in this specification.The compositions can also include any number of combinations ofadditional ingredients described throughout this specification (e.g.,pigments, or additional cosmetic or pharmaceutical ingredients). Theconcentrations of any ingredient within the compositions can vary. Innon-limiting embodiments, for example, the compositions can comprise,consist essentially of, or consist of, in their final form, for example,at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%,0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%,0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%,0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%,0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%,0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%,0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0.0053%, 0.0054%,0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%,0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%,0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%,0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%,0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%,0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%,0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%,0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%,0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%,0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%,0.1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%,0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000%, 0.5250%, 0.0550%,0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%,0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%, 0.9500%,0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%,2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%,3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%,4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%,5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%,6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%,8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%,9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 11%, 12%, 13%, 14%,15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%,29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or99% or any range derivable therein, of at least one of the ingredientsthat are mentioned throughout the specification and claims. Innon-limiting aspects, the percentage can be calculated by weight orvolume of the total composition. A person of ordinary skill in the artwould understand that the concentrations can vary depending on theaddition, substitution, and/or subtraction of ingredients in a givencomposition.

C. Vehicles

The compositions of the present invention can include or be incorporatedinto all types of vehicles and carriers. The vehicle or carrier can be apharmaceutically or dermatologically acceptable vehicle or carrier.Non-limiting examples of vehicles or carriers include water, glycerin,alcohol, oil, a silicon containing compound, a silicone compound, andwax. Variations and other appropriate vehicles will be apparent to theskilled artisan and are appropriate for use in the present invention. Incertain aspects, the concentrations and combinations of the compounds,ingredients, and agents can be selected in such a way that thecombinations are chemically compatible and do not form complexes whichprecipitate from the finished product.

D. Structure

The compositions of the present invention can be structured orformulated into a variety of different forms. Non-limiting examplesinclude emulsions (e.g., water-in-oil, water-in-oil-in-water,oil-in-water, silicone-in-water, water-in-silicone, oil-in-water-in-oil,oil-in-water-in-silicone emulsions), creams, lotions, solutions (bothaqueous and hydro-alcoholic), anhydrous bases (such as lipsticks andpowders), gels, masks, peels, and ointments. Variations and otherstructures will be apparent to the skilled artisan and are appropriatefor use in the present invention.

E. Additional Ingredients

In addition to the combination of ingredients disclosed by theinventors, the compositions can also include additional ingredients suchas cosmetic ingredients and pharmaceutical active ingredients.Non-limiting examples of these additional ingredients are described inthe following subsections.

1. Cosmetic Ingredients

The CTFA International Cosmetic Ingredient Dictionary and Handbook (2004and 2008) describes a wide variety of non-limiting cosmetic ingredientsthat can be used in the context of the present invention. Examples ofthese ingredient classes include: fragrance agents (artificial andnatural; e.g., gluconic acid, phenoxyethanol, and triethanolamine), dyesand color ingredients (e.g., Blue 1, Blue 1 Lake, Red 40, titaniumdioxide, D&C blue no. 4, D&C green no. 5, D&C orange no. 4, D&C red no.17, D&C red no. 33, D&C violet no. 2, D&C yellow no. 10, and D&C yellowno. 11), flavoring agents/aroma agents (e.g., Stevia rebaudiana(sweetleaf) extract, and menthol), adsorbents, lubricants, solvents,moisturizers (including, e.g., emollients, humectants, film formers,occlusive agents, and agents that affect the natural moisturizationmechanisms of the skin), water-repellants, UV absorbers (physical andchemical absorbers such as para-aminobenzoic acid (“PABA”) andcorresponding PABA derivatives, titanium dioxide, zinc oxide, etc.),essential oils, vitamins (e.g., A, B, C, D, E, and K), trace metals(e.g., zinc, calcium and selenium), anti-irritants (e.g., steroids andnon-steroidal anti-inflammatories), botanical extracts (e.g., Aloe vera,chamomile, cucumber extract, Ginkgo biloba, ginseng, and rosemary),anti-microbial agents, antioxidants (e.g., BHT and tocopherol),chelating agents (e.g., di sodium EDTA and tetrasodium EDTA),preservatives (e.g., methylparaben and propylparaben), pH adjusters(e.g., sodium hydroxide and citric acid), absorbents (e.g., aluminumstarch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrin,talc, and zeolite), skin bleaching and lightening agents (e.g.,hydroquinone and niacinamide lactate), humectants (e.g., sorbitol, urea,methyl gluceth-20, saccharide isomerate, and mannitol), exfoliants,waterproofing agents (e.g., magnesium/aluminum hydroxide stearate), skinconditioning agents (e.g., aloe extracts, allantoin, bisabolol,ceramides, dimethicone, hyaluronic acid, biosaccharide gum-1,ethylhexylglycerin, pentylene glycol, hydrogenated polydecene,octyldodecyl oleate, and dipotassium glycyrrhizate). Non-limitingexamples of some of these ingredients are provided in the followingsubsections.

a. UV Absorption and/or Reflecting Agents

UV absorption and/or reflecting agents that can be used in combinationwith the compositions of the present invention include chemical andphysical sunblocks. Non-limiting examples of chemical sunblocks that canbe used include para-aminobenzoic acid (PABA), PABA esters (glycerylPABA, amyldimethyl PABA and octyldimethyl PABA), butyl PABA, ethyl PABA,ethyl dihydroxypropyl PABA, benzophenones (oxybenzone, sulisobenzone,benzophenone, and benzophenone-1 through 12), cinnamates (octylmethoxycinnamate (octinoxate), isoamyl p-methoxycinnamate, octylmethoxycinnamate, cinoxate, diisopropyl methyl cinnamate, DEA-methoxycinnamate,ethyl diisopropylcinnamate, glyceryl octanoate dimethoxycinnamate andethyl methoxycinnamate), cinnamate esters, salicylates (homomethylsalicylate, benzyl salicylate, glycol salicylate, isopropylbenzylsalicylate, etc.), anthranilates, ethyl urocanate, homosalate,octisalate, dibenzoylmethane derivatives (e.g., avobenzone),octocrylene, octyl triazone, digalloyl trioleate, glycerylaminobenzoate, lawsone with dihydroxyacetone, ethylhexyl triazone,dioctyl butamido triazone, benzylidene malonate polysiloxane,terephthalylidene dicamphor sulfonic acid, disodium phenyldibenzimidazole tetrasulfonate, diethylamino hydroxybenzoyl hexylbenzoate, bis diethylamino hydroxybenzoyl benzoate, bisbenzoxazoylphenyl ethylhexylimino triazine, drometrizole trisiloxane,methylene bis-benzotriazolyl tetramethylbutylphenol, andbis-ethylhexyloxyphenol methoxyphenyltriazine, 4-methylbenzylidenecamphor, and isopentyl 4-methoxycinnamate. Non-limiting examples ofphysical sunblocks include, kaolin, talc, petrolatum and metal oxides(e.g., titanium dioxide and zinc oxide).

b. Moisturizing Agents

Non-limiting examples of moisturizing agents that can be used with thecompositions of the present invention include amino acids, chondroitinsulfate, diglycerin, erythritol, fructose, glucose, glycerin, glycerolpolymers, glycol, 1,2,6-hexanetriol, honey, hyaluronic acid,hydrogenated honey, hydrogenated starch hydrolysate, inositol, lactitol,maltitol, maltose, mannitol, natural moisturizing factor, PEG-15butanediol, polyglyceryl sorbitol, salts of pyrrolidone carboxylic acid,potassium PCA, propylene glycol, saccharide isomerate, sodiumglucuronate, sodium PCA, sorbitol, sucrose, trehalose, urea, andxylitol.

Other examples include acetylated lanolin, acetylated lanolin alcohol,threonine, lysine, alanine, algae extract, Aloe barbadensis, Aloebarbadensis extract, Aloe barbadensis gel, Althea officinalis extract,apricot (Prunus armeniaca) kernel oil, arginine, arginine aspartate,Arnica montana extract, aspartic acid, avocado (Persea gratissima) oil,barrier sphingolipids, butyl alcohol, beeswax, behenyl alcohol,beta-sitosterol, birch (Betula alba) bark extract, borage (Boragoofficinalis) extract, butcherbroom (Ruscus aculeatus) extract, butyleneglycol, Calendula officinalis extract, Calendula officinalis oil,candelilla (Euphorbia cerifera) wax, canola oil, caprylic/caprictriglyceride, cardamom (Elettaria cardamomum) oil, carnauba (Coperniciacerifera) wax, carrot (Daucus carota sativa) oil, castor (Ricinuscommunis) oil, ceramides, ceresin, ceteareth-5, ceteareth-12,ceteareth-20, cetearyl ethylhexanoate, ceteth-20, ceteth-24, cetylacetate, cetyl octanoate, cetyl palmitate, chamomile (Anthemis nobilis)oil, cholesterol, cholesterol esters, cholesteryl hydroxystearate,citric acid, clary (Salvia sclarea) oil, cocoa (Theobroma cacao) butter,coco-caprylate/caprate, coconut (Cocos nucifera) oil, collagen, collagenamino acids, corn (Zea mays) oil, fatty acids, decyl oleate, dimethiconecopolyol, dimethiconol, dioctyl adipate, dioctyl succinate,dipentaerythrityl hexacaprylate/hexacaprate, DNA, erythritol,ethoxydiglycol, ethyl linoleate, Eucalyptus globulus oil, eveningprimrose (Oenothera biennis) oil, fatty acids, Geranium maculatum oil,glucosamine, glucose glutamate, glutamic acid, glycereth-26, glycerin,glycerol, glyceryl distearate, glyceryl hydroxystearate, glyceryllaurate, glyceryl linoleate, glyceryl myristate, glyceryl oleate,glyceryl stearate, glyceryl stearate SE, glycine, glycol stearate,glycol stearate SE, glycosaminoglycans, grape (Vitis vinifera) seed oil,hazel (Corylus americana) nut oil, hazel (Corylus avellana) nut oil,hexylene glycol, hyaluronic acid, hybrid safflower (Carthamustinctorius) oil, hydrogenated castor oil, hydrogenated coco-glycerides,hydrogenated coconut oil, hydrogenated lanolin, hydrogenated lecithin,hydrogenated palm glyceride, hydrogenated palm kernel oil, hydrogenatedsoybean oil, hydrogenated tallow glyceride, hydrogenated vegetable oil,hydrolyzed collagen, hydrolyzed elastin, hydrolyzed glycosaminoglycans,hydrolyzed keratin, hydrolyzed soy protein, hydroxylated lanolin,proline, hydroxyproline, isocetyl stearate, isocetyl stearoyl stearate,isodecyl oleate, isopropyl isostearate, isopropyl lanol ate, isopropylmyristate, isopropyl palmitate, isopropyl stearate, isostearamide DEA,isostearic acid, isostearyl lactate, isostearyl neopentanoate, jasmine(Jasminum officinale) oil, jojoba (Buxus chinensis) oil, kelp, kukui(Aleurites moluccana) nut oil, lactamide MEA, laneth-16, laneth-10acetate, lanolin, lanolin acid, lanolin alcohol, lanolin oil, lanolinwax, lavender (Lavandula angustifolia) oil, lecithin, lemon (Citrusmedica limonum) oil, linoleic acid, linolenic acid, Macadamia ternifolianut oil, maltitol, matricaria (Chamomilla recutita) oil, methyl glucosesesquistearate, methyl silanol PCA, mineral oil, mink oil, mortierellaoil, myristyl lactate, myristyl myristate, myristyl propionate,neopentyl glycol dicaprylate/dicaprate, octyldodecanol, octyldodecylmyristate, octyldodecyl stearoyl stearate, octyl hydroxystearate, octylpalmitate, octyl salicylate, octyl stearate, oleic acid, olive (Oleaeuropaea) oil, orange (Citrus aurantium dulcis) oil, palm (Elaeisguineensis) oil, palmitic acid, pantethine, panthenol, panthenyl ethylether, paraffin, PCA, peach (Prunus persica) kernel oil, peanut (Arachishypogaea) oil, PEG-8 C12-18 ester, PEG-15 cocamine, PEG-150 distearate,PEG-60 glyceryl isostearate, PEG-5 glyceryl stearate, PEG-30 glycerylstearate, PEG-7 hydrogenated castor oil, PEG-40 hydrogenated castor oil,PEG-60 hydrogenated castor oil, PEG-20 methyl glucose sesquistearate,PEG-40 sorbitan peroleate, PEG-5 soy sterol, PEG-10 soy sterol, PEG-2stearate, PEG-8 stearate, PEG-20 stearate, PEG-32 stearate, PEG-40stearate, PEG-50 stearate, PEG-100 stearate, PEG-150 stearate,pentadecalactone, peppermint (Mentha piperita) oil, petrolatum,phospholipids, plankton extract, polyamino sugar condensate,polyglyceryl-3 diisostearate, polyquaternium-24, polysorbate 20,polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85,potassium myristate, potassium palmitate, propylene glycol, propyleneglycol dicaprylate/dicaprate, propylene glycol dioctanoate, propyleneglycol dipelargonate, propylene glycol laurate, propylene glycolstearate, propylene glycol stearate SE, PVP, pyridoxine dipalmitate,retinol, retinyl palmitate, rice (Oryza sativa) bran oil, RNA, rosemary(Rosmarinus officinalis) oil, rose oil, safflower (Carthamus tinctorius)oil, sage (Salvia officinalis) oil, sandalwood (Santalum album) oil,serine, serum protein, sesame (Sesamum indicum) oil, shea butter(Butyrospermum parkii), silk powder, sodium chondroitin sulfate, sodiumhyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodiumpolyglutamate, soluble collagen, sorbitan laurate, sorbitan oleate,sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol,soybean (Glycine soja) oil, sphingolipids, squalane, squalene,stearamide MEA-stearate, stearic acid, stearoxy dimethicone,stearoxytrim ethyl silane, stearyl alcohol, stearyl glycyrrhetinate,stearyl heptanoate, stearyl stearate, sunflower (Helianthus annuus) seedoil, sweet almond (Prunus amygdalus dulcis) oil, synthetic beeswax,tocopherol, tocopheryl acetate, tocopheryl linoleate, tribehenin,tridecyl neopentanoate, tridecyl stearate, triethanolamine, tristearin,urea, vegetable oil, water, waxes, wheat (Triticum vulgare) germ oil,and ylang (Cananga odorata) oil.

c. Antioxidants

Non-limiting examples of antioxidants that can be used with thecompositions of the present invention include acetyl cysteine, ascorbicacid polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanolpectinate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, cysteine, cysteine HCL, diamylhydroquinone,di-t-butylhydroquinone, dicetyl thiodipropionate, dioleyl tocopherylmethylsilanol, disodium ascorbyl sulfate, distearyl thiodipropionate,ditridecyl thiodipropionate, dodecyl gallate, erythorbic acid, esters ofascorbic acid, ethyl ferulate, ferulic acid, gallic acid esters,hydroquinone, isooctyl thioglycolate, kojic acid, magnesium ascorbate,magnesium ascorbyl phosphate, methylsilanol ascorbate, natural botanicalanti-oxidants such as green tea or grape seed extracts,nordihydroguaiaretic acid, octyl gallate, phenylthioglycolic acid,potassium ascorbyl tocopheryl phosphate, potassium sulfite, propylgallate, quinones, rosmarinic acid, sodium ascorbate, sodium bisulfite,sodium erythorbate, sodium metabisulfite, sodium sulfite, superoxidedismutase, sodium thioglycolate, sorbityl furfural, thiodiglycol,thiodiglycolamide, thiodiglycolic acid, thioglycolic acid, thiolacticacid, thiosalicylic acid, tocophereth-5, tocophereth-10, tocophereth-12,tocophereth-18, tocophereth-50, tocopherol, tocophersolan, tocopherylacetate, tocopheryl linoleate, tocopheryl nicotinate, tocopherylsuccinate, and tris(nonylphenyl)phosphite.

d. Structuring Agents

In other non-limiting aspects, the compositions of the present inventioncan include a structuring agent. Structuring agents, in certain aspects,assist in providing rheological characteristics to the composition tocontribute to the composition's stability. In other aspects, structuringagents can also function as an emulsifier or surfactant. Non-limitingexamples of structuring agents include stearic acid, palmitic acid,stearyl alcohol, cetyl alcohol, behenyl alcohol, stearic acid, palmiticacid, the polyethylene glycol ether of stearyl alcohol having an averageof about 1 to about 21 ethylene oxide units, the polyethylene glycolether of cetyl alcohol having an average of about 1 to about 5 ethyleneoxide units, and mixtures thereof.

e. Emulsifiers

In certain aspects of the present invention, the compositions do notinclude an emulsifier. In other aspects, however, the compositions caninclude one or more emulsifiers. Emulsifiers can reduce the interfacialtension between phases and improve the formulation and stability of anemulsion. The emulsifiers can be nonionic, cationic, anionic, andzwitterionic emulsifiers (See McCutcheon' s (1986); U.S. Pat. Nos.5,011,681; 4,421,769; 3,755,560). Non-limiting examples include estersof glycerin, esters of propylene glycol, fatty acid esters ofpolyethylene glycol, fatty acid esters of polypropylene glycol, estersof sorbitol, esters of sorbitan anhydrides, carboxylic acid copolymers,esters and ethers of glucose, ethoxylated ethers, ethoxylated alcohols,alkyl phosphates, polyoxyethylene fatty ether phosphates, fatty acidamides, acyl lactylates, soaps, TEA stearate, DEA oleth-3 phosphate,polyethylene glycol 20 sorbitan monolaurate (polysorbate 20),polyethylene glycol 5 soya sterol, steareth-2, steareth-20, steareth-21,ceteareth-20, cetearyl glucoside, cetearyl alcohol, C12-13 pareth-3,PPG-2 methyl glucose ether distearate, PPG-5-ceteth-20,bis-PEG/PPG-20/20 dimethicone, ceteth-10, polysorbate 80, cetylphosphate, potassium cetyl phosphate, diethanolamine cetyl phosphate,polysorbate 60, glyceryl stearate, PEG-100 stearate, arachidyl alcohol,arachidyl glucoside, hydroxypropyl cyclodextrin, and mixtures thereof.

f. Silicone Containing Compounds

In non-limiting aspects, silicone containing compounds include anymember of a family of polymeric products whose molecular backbone ismade up of alternating silicon and oxygen atoms with side groupsattached to the silicon atoms. By varying the —Si—O— chain lengths, sidegroups, and crosslinking, silicones can be synthesized into a widevariety of materials. They can vary in consistency from liquid to gel tosolids.

The silicone containing compounds that can be used in the context of thepresent invention include those described in this specification or thoseknown to a person of ordinary skill in the art. Non-limiting examplesinclude silicone oils (e.g., volatile and non-volatile oils), gels, andsolids. In certain aspects, the silicon containing compounds includesilicone oils such as a polyorganosiloxane. Non-limiting examples ofpolyorganosiloxanes include dimethicone, cyclomethicone,cyclohexasiloxane, poly silicone-11, phenyl trimethicone,trimethylsilylamodimethicone, stearoxytrimethylsilane, or mixtures ofthese and other organosiloxane materials in any given ratio in order toachieve the desired consistency and application characteristicsdepending upon the intended application (e.g., to a particular area suchas the skin, hair, or eyes). A “volatile silicone oil” includes asilicone oil have a low heat of vaporization, i.e. normally less thanabout 50 cal per gram of silicone oil. Non-limiting examples of volatilesilicone oils include: cyclomethicones such as Dow Corning 344 Fluid,Dow Corning 345 Fluid, Dow Corning 244 Fluid, and Dow Corning 245 Fluid,Volatile Silicon 7207 (Union Carbide Corp., Danbury, Conn.); lowviscosity dimethicones, i.e. dimethicones having a viscosity of about 50cst or less (e.g., dimethicones such as Dow Corning 200-0.5 cst Fluid).The Dow Corning Fluids are available from Dow Corning Corporation,Midland, Mich. Cyclomethicone and dimethicone are described in the ThirdEdition of the CTFA Cosmetic Ingredient Dictionary (incorporated byreference) as cyclic dimethyl polysiloxane compounds and a mixture offully methylated linear siloxane polymers end-blocked withtrimethylsiloxy units, respectively. Other non-limiting volatilesilicone oils that can be used in the context of the present inventioninclude those available from General Electric Co., Silicone ProductsDiv., Waterford, N.Y. and SWS Silicones Div. of Stauffer Chemical Co.,Adrian, Mich.

g. Exfoliating Agent

Exfoliating agents include ingredients that remove dead skin cells onthe skin's outer surface. These agents may act through mechanical,chemical, and/or other means. Non-limiting examples of mechanicalexfoliating agents include abrasives such as pumice, silica, cloth,paper, shells, beads, solid crystals, solid polymers, etc. Non-limitingexamples of chemical exfoliating agents include acids and enzymeexfoliants. Acids that can be used as exfoliating agents include, butare not limited to, glycolic acid, lactic acid, citric acid, alphahydroxy acids, beta hydroxy acids, etc. Other exfoliating agents knownto those of skill in the art are also contemplated as being usefulwithin the context of the present invention.

h. Essential Oils

Essential oils include oils derived from herbs, flowers, trees, andother plants. Such oils are typically present as tiny droplets betweenthe plant's cells, and can be extracted by several method known to thoseof skill in the art (e.g., steam distilled, enfleurage (i.e., extractionby using fat), maceration, solvent extraction, or mechanical pressing).When these types of oils are exposed to air they tend to evaporate(i.e., a volatile oil). As a result, many essential oils are colorless,but with age they can oxidize and become darker. Essential oils areinsoluble in water and are soluble in alcohol, ether, fixed oils(vegetal), and other organic solvents. Typical physical characteristicsfound in essential oils include boiling points that vary from about 160°to 240° C. and densities ranging from about 0.759 to about 1.096.

Essential oils typically are named by the plant from which the oil isfound. For example, rose oil or peppermint oil are derived from rose orpeppermint plants, respectively. Non-limiting examples of essential oilsthat can be used in the context of the present invention include sesameoil, macadamia nut oil, tea tree oil, evening primrose oil, Spanish sageoil, Spanish rosemary oil, coriander oil, thyme oil, pimento berriesoil, rose oil, anise oil, balsam oil, bergamot oil, rosewood oil, cedaroil, chamomile oil, sage oil, clary sage oil, clove oil, cypress oil,eucalyptus oil, fennel oil, sea fennel oil, frankincense oil, geraniumoil, ginger oil, grapefruit oil, jasmine oil, juniper oil, lavender oil,lemon oil, lemongrass oil, lime oil, mandarin oil, marjoram oil, myrrhoil, neroli oil, orange oil, patchouli oil, pepper oil, black pepperoil, petitgrain oil, pine oil, rose otto oil, rosemary oil, sandalwoodoil, spearmint oil, spikenard oil, vetiver oil, wintergreen oil, orylang. Other essential oils known to those of skill in the art are alsocontemplated as being useful within the context of the presentinvention.

i. Thickening Agents

Thickening agents, including thickener or gelling agents, includesubstances which that can increase the viscosity of a composition.Thickeners includes those that can increase the viscosity of acomposition without substantially modifying the efficacy of the activeingredient within the composition. Thickeners can also increase thestability of the compositions of the present invention. In certainaspects of the present invention, thickeners include hydrogenatedpolyisobutene, trihydroxystearin, ammonium acryloyldimethyltaurate/vpcopolymer, or a mixture of them.

Non-limiting examples of additional thickening agents that can be usedin the context of the present invention include carboxylic acidpolymers, crosslinked polyacrylate polymers, polyacrylamide polymers,polysaccharides, and gums. Examples of carboxylic acid polymers includecrosslinked compounds containing one or more monomers derived fromacrylic acid, substituted acrylic acids, and salts and esters of theseacrylic acids and the substituted acrylic acids, wherein thecrosslinking agent contains two or more carbon-carbon double bonds andis derived from a polyhydric alcohol (see U.S. Pat. Nos. 5,087,445;4,509,949; 2,798,053; CTFA International Cosmetic Ingredient Dictionary,Fourth edition, 1991, pp. 12 and 80). Examples of commercially availablecarboxylic acid polymers include carbomers, which are homopolymers ofacrylic acid crosslinked with allyl ethers of sucrose or pentaerythritol(e.g., CARBOPOL™ 900 series from B. F. Goodrich).

Non-limiting examples of crosslinked polyacrylate polymers includecationic and nonionic polymers. Examples are described in U.S. Pat. Nos.5,100,660 ; 4,849,484; 4,835,206; 4,628,078; 4,599,379).

Non-limiting examples of polyacrylamide polymers (including nonionicpolyacrylamide polymers including substituted branched or unbranchedpolymers) include polyacrylamide, isoparaffin and laureth-7, multi-blockcopolymers of acrylamides and substituted acrylamides with acrylic acidsand substituted acrylic acids.

Non-limiting examples of polysaccharides include cellulose,carboxymethyl hydroxyethylcellulose, cellulose acetate propionatecarboxylate, hydroxyethylcellulose, hydroxyethyl ethylcellulose,hydroxypropylcellulose, hydroxypropyl methylcellulose, methylhydroxyethylcellulose, microcrystalline cellulose, sodium cellulosesulfate, and mixtures thereof. Another example is an alkyl substitutedcellulose where the hydroxy groups of the cellulose polymer ishydroxyalkylated (preferably hydroxy ethylated or hydroxypropylated) toform a hydroxyalkylated cellulose which is then further modified with aC10 -C30 straight chain or branched chain alkyl group through an etherlinkage. Typically these polymers are ethers of C10-C30 straight orbranched chain alcohols with hydroxyalkylcelluloses. Other usefulpolysaccharides include scleroglucans comprising a linear chain of (1-3)linked glucose units with a (1-6) linked glucose every three unit.

Non-limiting examples of gums that can be used with the presentinvention include acacia, agar, algin, alginic acid, ammonium alginate,amylopectin, calcium alginate, calcium carrageenan, carnitine,carrageenan, dextrin, gelatin, gellan gum, guar gum, guarhydroxypropyltrimonium chloride, hectorite, hyaluronic acid, hydratedsilica, hydroxypropyl chitosan, hydroxypropyl guar, karaya gum, kelp,locust bean gum, natto gum, potassium alginate, potassium carrageenan,propylene glycol alginate, sclerotium gum, sodium carboxymethyl dextran,sodium carrageenan, tragacanth gum, xanthan gum, and mixtures thereof

j. Preservatives

Non-limiting examples of preservatives that can be used in the contextof the present invention include quaternary ammonium preservatives suchas polyquaternium-1 and benzalkonium halides (e.g., benzalkoniumchloride (“BAC”) and benzalkonium bromide), parabens (e.g.,methylparabens and propylparabens), phenoxyethanol, benzyl alcohol,chlorobutanol, phenol, sorbic acid, thimerosal or combinations thereof.

2. Pharmaceutical Ingredients

Pharmaceutical active agents are also contemplated as being useful withthe compositions of the present invention. Non-limiting examples ofpharmaceutical active agents include anti-acne agents, agents used totreat rosacea, analgesics, anesthetics, anorectals, antihistamines,anti-inflammatory agents including non-steroidal anti-inflammatorydrugs, antibiotics, antifungals, antivirals, antimicrobials, anti-canceractives, scabicides, pediculicides, antineoplastics, antiperspirants,antipruritics, antipsoriatic agents, antiseborrheic agents, biologicallyactive proteins and peptides, burn treatment agents, cauterizing agents,depigmenting agents, depilatories, diaper rash treatment agents,enzymes, hair growth stimulants, hair growth retardants including DFMOand its salts and analogs, hemostatics, kerotolytics, canker soretreatment agents, cold sore treatment agents, dental and periodontaltreatment agents, photosensitizing actives, skin protectant/barrieragents, steroids including hormones and corticosteroids, sunburntreatment agents, sunscreens, transdermal actives, nasal actives,vaginal actives, wart treatment agents, wound treatment agents, woundhealing agents, etc.

F. Kits

Kits are also contemplated as being used in certain aspects of thepresent invention. For instance, compositions of the present inventioncan be included in a kit. A kit can include a container. Containers caninclude a bottle, a metal tube, a laminate tube, a plastic tube, adispenser, a pressurized container, a barrier container, a package, acompartment, a lipstick container, a compact container, cosmetic pansthat can hold cosmetic compositions, or other types of containers suchas injection or blow-molded plastic containers into which thedispersions or compositions or desired bottles, dispensers, or packagesare retained. The kit and/or container can include indicia on itssurface. The indicia, for example, can be a word, a phrase, anabbreviation, a picture, or a symbol.

The containers can dispense a pre-determined amount of the composition.In other embodiments, the container can be squeezed (e.g., metal,laminate, or plastic tube) to dispense a desired amount of thecomposition. The composition can be dispensed as a spray, an aerosol, aliquid, a fluid, or a semi-solid. The containers can have spray, pump,or squeeze mechanisms. A kit can also include instructions for employingthe kit components as well the use of any other compositions included inthe container. Instructions can include an explanation of how to apply,use, and maintain the compositions.

G. EXAMPLES

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

All of the compositions and methods disclosed and claimed herein can bemade and executed without undue experimentation in light of the presentdisclosure. While the compositions and methods of this invention havebeen described in terms of preferred embodiments, it will be apparent tothose of skill in the art that variations may be applied to thecompositions and methods and in the steps or in the sequence of steps ofthe method described herein without departing from the concept, spirit,and scope of the invention. More specifically, it will be apparent thatcertain agents which are both chemically and physiologically related maybe substituted for the agents described herein while the same or similarresults would be achieved. All such similar substitutes andmodifications apparent to those skilled in the art are deemed to bewithin the spirit, scope and concept of the invention as defined by theappended claims.

1. Example 1 (In Vitro Myotube Contraction Data)

Each of Rosmarinus officinalis leaf extract, Lavendula stoechas extract,and Acmella oleracea extract were found to reduce contraction ofmyotubes, which can result in reduced muscle contraction of a facialmuscle and reduced appearance of facial deep lines or wrinkles anddynamic facial wrinkles and forehead wrinkles that are caused due torepetitive facial muscle contraction. The methodology used to obtain thedata included: (1) Primary Human Skeletal muscle myoblasts (HSMM) weremade to differentiate into Myotubes; (2) Myotubes were then treated with‘inducers’ (Acetcholine), which in turn causes an influx of calcium andcreating an action potential; and (3) This action potential causes themuscle cells to contract. The concentration amounts of each extract usedand the resulting effect on inhibition of calcium levels are provided inTable 1.

TABLE 1 Ingredient Inhibition of Calcium Levels Rosmarinus OfficinalisLeaf extract* 1.0 wt. % conc. = 100% inhibition 2.0 wt. % conc. = 100%inhibition Lavendula Stoechas Extract** 0.5% wt. conc. = 30% inhibition1.0% wt. conc. = 30% inhibition Acmella Oleracea Extract*** 0.5% wt.conc. = 77% inhibition 1.0% wt. conc. = 96% inhibition *Rosmarinusofficinalis leaf extract was a eutectic extract in which the solventused to prepare the extract was a fluid mixture of betaine, lactic acid,and water. The extract was supplied by Naturex (France) under the tradename ROSEMARY EUTECTYS BLA ™. **Lavendula stoechas extract used was asupercritical CO₂ extract of the flower/leaf/stem of Lavendula stoechas.The CO₂ extract was mixed with caprylic/capric triglycerides. Theextract was supplied by Barnet Products LLC (Englewood Cliffs, NewJersey (USA)) under the trade name STOCHEY'S. ***Acmella oleraceaextract used was a hydro-ethanolic extract in which the solvent includeda fluid mixture of water, ethanol, and 1,3 propanediol. The ethanol wasremoved from the resulting extraction fluid. The extract was supplied byGattefossé (France) under the trade name GATULINE ® EXPRESSION AF.

2. Example 2 (In Vitro Collagen Expression Data)

Acmella oleracea extract was found to increase collagen production inhuman dermal fibroblasts. The data and amount of extract used isprovided in Table 2.

TABLE 2 Ingredient Increased Collagen Production Acmella OleraceaExtract* 1.0 wt. % conc. = 61% increased collagen production *Acmellaoleracea extract used was a hydro-ethanolic extract in which the solventincluded a fluid mixture of water, ethanol, and 1,3 propanediol. Theethanol was removed from the resulting extraction fluid. The extract wassupplied by Gattefossé (France) under the trade name GATULINE ®EXPRESSION AF.

Collagen is an extracellular matrix protein critical for skin structure.Increased synthesis of collagen helps improve skin firmness andelasticity. This bioassay was used to examine the effect of Acmellaoleracea extract on the production of procollagen peptide (a precursorto collagen) by human dermal fibroblasts. The endpoint of this assay wasa spectrophotometric measurement that reflected the presence ofprocollagen peptide and cellular viability. The assay employed thequantitative sandwich enzyme immunoassay technique whereby a monoclonalantibody specific for procollagen peptide was pre-coated onto amicroplate. Standards and samples were pipetted into the wells and anyprocollagen peptide present was bound by the immobilized antibody. Afterwashing away any unbound substances, an enzyme-linked polyclonalantibody specific for procollagen peptide was added to the wells.Following a wash to remove any unbound antibody-enzyme reagent, asubstrate solution was added to the wells and color developed inproportion to the amount of procollagen peptide bound in the initialstep. Color development was stopped and the intensity of the color at450 nm was measured using a microplate reader.

For generation of samples and controls, subconfluent normal human adultdermal fibroblasts (Cascade Biologics) were cultivated in standard DMEMgrowth medium with 10% fetal bovine serum (Mediatech) at 37° C. in 10%CO₂. The cells were treated with each of the tested ingredients andcontrols for 3 days. Following incubation, cell culture medium wascollected and the amount of procollagen peptide secretion was quantifiedusing the sandwich enzyme linked immuno-sorbant assay (ELISA) fromTakara (#MK101) as explained above.

3. Example 3 (In Vitro Laminin Expression Data)

Acmella oleracea extract was found to increase laminin production inhuman dermal fibroblasts. The data and amount of extract used isprovided in Table 3.

TABLE 3 Ingredient Increased Laminin Production Acmella OleraceaExtract* 1.0 wt. % conc. = 53% increased laminin production *Acmellaoleracea extract used was a hydro-ethanolic extract in which the solventincluded a fluid mixture of water, ethanol, and 1,3 propanediol. Theethanol was removed from the resulting extraction fluid. The extract wassupplied by Gattefossé (France) under the trade name GATULINE ®EXPRESSION AF.

Laminin is a major protein in the dermal-epidermal junction (DEJ) (alsoreferred to as the basement membrane). The DEJ is located between thedermis and the epidermis and interlocks forming fingerlike projectionscalled rete ridges. The cells of the epidermis receive their nutrientsfrom the blood vessels in the dermis. The rete ridges increase thesurface area of the epidermis that is exposed to these blood vessels andthe needed nutrients. The DEJ provides adhesion of the two tissuecompartments and governs the structural integrity of the skin. Lamininis a structural glycoproteins located in the DEJ. Considered a glue thatholds the cells together, laminin is secreted by dermal fibroblasts tohelp facilitate intra- and inter-cellular adhesion of the epidermalcells to the DEJ. Laminin secretion was monitored by quantifying lamininin cell supernatants of cultured human fibroblasts treated for 3 dayswith culture medium with the test ingredient (Acmella oleracea extract).Following incubation, laminin was measured using immunofluorescentantibodies directed against each protein in an enzyme linkedimmuno-sorbant assay (ELISA). Measurements were normalized for cellularmetabolic activity, as determined by bioconversion of3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS).

4. Example 4 (In Vitro MMP Inhibition Data)

Rosmarinus officinalis leaf extract was found to inhibit matrixmetalloproteinases (MMP) 1, 3, and 9 in human dermal fibroblasts. Thedata and amount of extract used is provided in Table 4.

TABLE 4 Ingredient Inhibition of MMP-1, 3, and 9 Production RosmarinusOfficinalis MMP-1 @1.0 wt. % conc. = −98% inhibition Leaf extract* MMP-3@1.0 wt. % conc. = −39% inhibition MMP-9 @1.0 wt. % conc. = −61%inhibition *Rosmarinus officinalis leaf extract was a eutectic extractin which the solvent used to prepare the extract was fluid mixture ofbetaine, lactic acid, and water, The extract was supplied by Naturex(France) under the trade name ROSEMARY EUTECTYS BLA ™.

Matrix Metalloproteinase 1 Enzyme Activity (MMP-1) Assay: MMPs areextracellular proteases that play a role in many normal and diseasestates by virtue of their broad substrate specificity. MMP-1 substratesinclude collagen IV. The Molecular Probes Enz/ChekGelatinase/Collagenase Assay kit (#E12055) utilizes a fluorogenicgelatin substrate to detect MMP1 protease activity. Upon proteolyticcleavage, bright green fluorescence is revealed and may be monitoredusing a fluorescent microplate reader to measure enzymatic activity. TheEnz/Chek Gelatinase/Collagenase Assay kit (#E12055) from Invitrogen isdesigned as an in vitro assay to measure MMP-1 enzymatic activity. TheRosmarinus officinalis leaf extract was assayed. The assay relies uponthe ability of purified MMP-1 enzyme to degrade a fluorogenic gelatinsubstrate. Once the substrate is specifically cleaved by MMP-1 brightgreen fluorescence is revealed and was monitored using a fluorescentmicroplate reader. Rosmarinus officinalis leaf extract was incubated inthe presence or absence of the purified enzyme and substrate todetermine their protease inhibitor capacity.

Matrix Metalloproteinase 3 and 9 Enzyme Activity (MMP-3 and MMP-9)Assay: MMP-3 substrates include collagens, fibronectins, and laminin;while MMP-9 substrates include collagen VII, fibronectins and laminin.Using Colorimetric Drug Discovery kits from BioMol International forMMP3 (AK-400) and MMP-9 (AK-410), this assay was designed to measureprotease activity of MMPs using a thiopeptide as a chromogenic substrate(Ac-PLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5)5,6. The MMP cleavagesite peptide bond is replaced by a thioester bond in the thiopeptide.Hydrolysis of this bond by an MMP produces a sulfhydryl group, whichreacts with DTNB [5,5′-dithiobis(2-nitrobenzoic acid), Ellman's reagent]to form 2-nitro-5-thiobenzoic acid, which can be detected by itsabsorbance at 412 nm (ε=13,600 M-1 cm-1 at pH 6.0 and above 7). TheRosmarinus officinalis leaf extract was assayed.

5. Example 5 (In Vivo Clinical Data)

In vivo clinical data on the combination of Rosmarinus officinalis leafextract, Lavendula stoechas extract, and Acmella oleracea extract wasobtained. The Rosmarinus officinalis leaf extract was a eutectic extractin which the solvent used to prepare the extract was a fluid mixture ofbetaine, lactic acid, and water. The extract was supplied by Naturex(France) under the trade name ROSEMARY EUTECTYS BLA™. The Lavendulastoechas extract used was a supercritical CO₂ extract of theflower/leaf/stem of Lavendula stoechas. The CO₂ extract was mixed withcaprylic/capric triglycerides. The extract was supplied by BarnetProducts LLC (Englewood Cliffs, N.J. (USA)) under the trade nameSTOCHEY'S. The Acmella oleracea extract used was a hydro-ethanolicextract in which the solvent included a fluid mixture of water, ethanol,and 1,3 propanediol. The ethanol was removed from the resultingextraction fluid. The extract was supplied by Gattefosse (France) underthe trade name GATULINE® EXPRESSION AF.

The purpose of the in vivo clinical study was to evaluate the efficacyof a combination of the three extracts for muscle relaxation and wrinklereduction over four weeks of twice-daily application on 30 individuals(n=30). The combination of the three extracts was placed in adermatologically acceptable vehicle. The dermatologically acceptablevehicle was designed to test the efficacy of the combination of thethree extracts. The other ingredients in the dermatologically acceptablevehicle were not believed to contribute to the clinical data results ineither a positive or negative manner. The amounts of each extract usedin the dermatologically acceptable vehicle was Rosmarinus officinalisleaf extract at 1 wt. %, Lavendula stoechas extract at 2 wt. %, andAcmella oleracea extract at 2 wt. %.

The in vivo clinical study design was a randomized controlled,investigator-blinded clinical study. The following parameters were used:(1) subjects—females between the ages of 35 and 70 in good generalhealth (no physical required) and were clinically determined to havescored 3-7 (inclusively) on a 10-point scale for moderatefrown/glabellar lines; (2) test site—glabella area;

(3) evaluation time points—baseline (week 0), week 2, week 4, and week8; (4) testing method—electromyography (EMG) for muscle response(mV/s):AUC (area under the curve); and (5) clinical endpointsmeasured—fine lines and wrinkles and skin texture on a 0 to 9 scale.

FIGS. 1-2 provide data illustrating muscle contraction inhibition viaEMG testing. FIGS. 3-4 provide data illustrating reduction in theappearance of fine lines and wrinkles. As illustrated in these FIGS. 1-4, the data suggests that the combination of Rosmarinus officinalis leafextract, Lavendula stoechas extract, and Acmella oleracea extract canreduce facial muscle contraction and can reduce the overall appearanceof fine lines and wrinkles. The following Table 5 provides dataillustrating the EMG response and clinical comparison between awell-known injectable neuromodulator (Botox® (onabotulinumtoxinA)) andthe combination of Rosmarinus officinalis leaf extract, Lavendulastoechas extract, and Acmella oleracea extract.

TABLE 5 % Clinical Improvement % EMG Response (AUC) of WrinklesCombination of Combination of BoTox - 5U three extracts BoTox - 5U threeextracts (n = 5)** (n = 30) (n = 5)** (n = 30) Week 2 58%* 11% 31%* 1%Week 4 40%* 10% 35%*  9%* Week 8 Not  16%* Not 19%* measured measured *p< 0.05 statistically significant improvement when compared to baseline.**The BoTox - 5U study was used as a control to validate the method andperformed on 5 individuals. Each individual was administered 5 units (U)of Botox to the glabella region and % EMG Response (AUC) was measured.This was done to standardize the aforementioned combination of extractsstudy.

6. Example 6 (In Vivo Clinical Data)

In a comparative clinical study, additional in vivo clinical data on thecombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract was obtained. The Rosmarinusofficinalis leaf extract was a eutectic extract in which the solventused to prepare the extract was a fluid mixture of betaine, lactic acid,and water. The extract was supplied by Naturex (France) under the tradename ROSEMARY EUTECTYS BLA™. The Lavendula stoechas extract used was asupercritical CO₂ extract of the flower/leaf/stem of Lavendula stoechas.The CO₂ extract was mixed with caprylic/capric triglycerides. Theextract was supplied by Barnet Products LLC (Englewood Cliffs, N.J.(USA)) under the trade name STOCHEY'S. The Acmella oleracea extract usedwas a hydro-ethanolic extract in which the solvent included a fluidmixture of water, ethanol, and 1,3 propanediol. The ethanol was removedfrom the resulting extraction fluid. The extract was supplied byGattefosse (France) under the trade name GATULINE® EXPRESSION AF.

The purpose of the comparative in vivo clinical study was to evaluatethe efficacy of a combination of the three extracts for musclerelaxation and wrinkle reduction over four weeks of twice-dailyapplication on Experimental Group 1 participants using a micro-roller(n=33) versus application on Experimental Group 2 participants withoutusing a micro-roller (n=30). The combination of the three extracts wasplaced in a dermatologically acceptable vehicle as the test product. Thedermatologically acceptable vehicle was designed to test the efficacy ofthe combination of the three extracts when applied with a micro-rollerversus application without a micro-roller. Study participants inExperimental Group 1 used a 1.0 mm microroller. Study participantsapplied the composition containing the three extracts between theireyebrows in the morning and the evening after cleansing their face witha personal cleanser. For Experimental Group 1, study participants usedthe micro-roller 3 times per week in the evenings by rolling themicro-roller back and forth 4 to 5 times between eyebrows changingdirections with each pass. For both Experimental Group 1 andExperimental Group 2, an occlusive patch (e.g., SCARAWAY®) was placedbetween eyebrows overnight. The other ingredients in thedermatologically acceptable vehicle were not believed to contribute tothe clinical data results in either a positive or negative manner. Theamounts of each extract used in the dermatologically acceptable vehiclewas Rosmarinus officinalis leaf extract at 1 wt. %, Lavendula stoechasextract at 2 wt. %, and Acmella oleracea extract at 2 wt. %.

The comparative in vivo clinical study design was a randomizedcontrolled, investigator-blinded clinical study. The followingparameters were used: (1) subjects—females between the ages of 35 and 70in good general health (no physical required) and were clinicallydetermined to have scored 3-7 (inclusively) on a 10-point scale formoderate frown/glabellar lines; (2) test site—glabella area; (3) testproduct—Frontalis; supplemental products—SCARAWAY® occlusive patch, andmicroroller; (4) evaluation time points—baseline (week 0), week 2, week4, and week 8; (5) testing method—electromyography (EMG) for muscleresponse (mV/s): AUC (area under the curve); and (6) clinical endpointsmeasured—fine lines and wrinkles and skin texture on a 0 to 9 scale.

FIGS. 5-6 provide data illustrating muscle contraction inhibition viaEMG testing comparing percent improvement between application with amicro-roller and application without a micro-roller. FIGS. 7-8 providedata illustrating reduction in the appearance of fine lines and wrinklescomparing percent improvement between application with a micro-rollerand application without a micro-roller. As illustrated in FIGS. 5-8 ,the data suggests that application of the composition containing thecombination of Rosmarinus officinalis leaf extract, Lavendula stoechasextract, and Acmella oleracea extract, combined with using amicro-roller to promote penetration of the composition into the skin,can reduce facial muscle contraction and can reduce the overallappearance of fine lines and wrinkles. It was determined thatapplication of the composition with a micro-roller was effective atattaining a faster and stronger reduction in facial muscle contractionat 2, 4, and 8 weeks compared to baseline, and reduced fine lines andwrinkles and skin texture within 4 weeks, where 79% of subjects showedimprovement in both fine lines and wrinkles and skin smoothness after 8weeks. It was further determined that application of the compositionwithout a micro-roller was still effective at attaining a reduction infacial muscle contraction after 8 weeks, and reduced fine lines andwrinkles and skin texture within 4 weeks, where 87% of subjects showedimprovement in both fine lines and wrinkles and 90% in skin smoothnessafter 8 weeks.

FIGS. 9-10 provide data illustrating improvement in skintexture/roughness comparing percent improvement between application witha micro-roller and application without a micro-roller. As illustrated inFIGS. 9-10 , the data suggests that application of the compositioncontaining the combination of Rosmarinus officinalis leaf extract,Lavendula stoechas extract, and Acmella oleracea extract, combined withusing a micro-roller to promote penetration of the composition into theskin, can significantly improve skin texture and roughness in theglabellar area after 2, 4, and 8 weeks. It was determined that 79% ofsubjects showed improvement in skin smoothness after 8 weeks afterapplication of the composition with a micro-roller. It was furtherdetermined that application of the composition without a micro-rollerwas still effective at improving skin smoothness after 8 weeks.

The following Table 6 provides data illustrating the EMG response andclinical comparison between a well-known injectable neuromodulator(Botox (onabotulinumtoxinA)) and the combination of Rosmarinusofficinalis leaf extract, Lavendula stoechas extract, and Acmellaoleracea extract applied with a micro-roller and applied without amicro-roller.

TABLE 6 % EMG Response (AUC) % Clinical Improvement of WrinklesExperimental Experimental Experimental Experimental Group 1 (w/ Group 2(w/o Group 1 (w/ Group 2 (w/o BoTox - 5 U micro-roller) micro-roller)BoTox - 5 U micro-roller) micro-roller) (n = 5)** (n = 33) (n = 30) (n =5)** (n = 33) (n = 30) Week 2 58%* 16%* 11% 31%* 3% 1% Week 4 40%* 16%*10% 35%*  9%*  9%* Week 8 Not 21%*  16%* Not 18%* 19%* measured measured*p < 0.05 statistically significant improvement when compared tobaseline. **The BoTox - 5 U study was used as a positive control incomparing Experimental Group 1 and Experimental Group 2 and wasperformed on 5 individuals. Each individual was administered 5 units (U)of Botox to the glabella region and % EMG Response (AUC) was measured.This was done to standardize the aforementioned combination of extractsstudy.

7. Example 7 (Additional Assays)

Assays that can be used to determine the efficacy of any one of theingredients or any combination of ingredients or compositions havingsaid combination of ingredients disclosed throughout the specificationand claims can be determined by methods known to those of ordinary skillin the art. The following are non-limiting assays that can be used inthe context of the present invention. It should be recognized that othertesting procedures can be used, including, for example, objective andsubjective procedures.

Antioxidant (AO) Assay: An in vitro bioassay that measures the totalanti-oxidant capacity of any one of the ingredients, combination ofingredients, or compositions having said combinations disclosed in thespecification. The assay relies on the ability of antioxidants in thesample to inhibit the oxidation of ABTS®(2,2′-azino-di-[3-ethylbenzthiazoline sulphonate]) to ABTS®⋅+bymetmyoglobin. The antioxidant system of living organisms includesenzymes such as superoxide dismutase, catalase, and glutathioneperoxidase; macromolecules such as albumin, ceruloplasmin, and ferritin;and an array of small molecules, including ascorbic acid, a-tocopherol,(3-carotene, reduced glutathione, uric acid, and bilirubin. The sum ofendogenous and food-derived antioxidants represents the totalantioxidant activity of the extracellular fluid. Cooperation of all thedifferent antioxidants provides greater protection against attack byreactive oxygen or nitrogen radicals, than any single compound alone.Thus, the overall antioxidant capacity may give more relevant biologicalinformation compared to that obtained by the measurement of individualcomponents, as it considers the cumulative effect of all antioxidantspresent in plasma and body fluids. The capacity of the antioxidants inthe sample to prevent ABTS oxidation is compared with that of Trolox, awater-soluble tocopherol analogue, and is quantified as molar Troloxequivalents. Anti-Oxidant capacity kit # 709001 from Cayman Chemical(Ann Arbor, Mich. USA) can be used as an in vitro bioassay to measurethe total anti-oxidant capacity of each of any one of the activeingredients, combination of ingredients, or compositions having saidcombinations disclosed in the specification. The protocol can befollowed according to manufacturer recommendations.

ORAC Assay: Oxygen Radical Absorption (or Absorbance) Capacity (ORAC) ofany one of the active ingredients, combination of ingredients, orcompositions having said combinations disclosed in the specification canalso be assayed by measuring the antioxidant activity of suchingredients or compositions. Antioxidant activity indicates a capabilityto reduce oxidizing agents (oxidants). This assay quantifies the degreeand length of time it takes to inhibit the action of an oxidizing agent,such as oxygen radicals, that are known to cause damage to cells (e.g.,skin cells). The ORAC value of any one of the active ingredients,combination of ingredients, or compositions having said combinationsdisclosed in the specification can be determined by methods known tothose of ordinary skill in the art (see U.S. Publication Nos.2004/0109905 and 2005/0163880; and commercially available kits such asZen-Bio ORAC Anti-oxidant Assay kit (#AOX-2)). The Zen-Bio ORACAnti-oxidant Assay kit measures the loss of fluorescein fluorescenceover time due to the peroxyl-radical formation by the breakdown of AAPH(2,2′-axobis-2-methyl propanimidamide, dihydrochloride). Trolox, a watersoluble vitamin E analog, serves as positive control inhibitionfluorescein decay in a dose dependent manner.

Cyclooxygenase (COX) Assay: An in vitro cyclooxygenase-1 and -2 (COX-1,-2) inhibition assay. COX is a bifunctional enzyme exhibiting bothcyclooxygenase and peroxidase activities. The cyclooxygenase activityconverts arachidonic acid to a hydroperoxy endoperoxide (ProstaglandinG2; PGG2) and the peroxidase component reduces the endoperoxide(Prostaglandin H2; PGH2) to the corresponding alcohol, the precursor ofprostaglandins, thromboxanes, and prostacyclins. This COX Inhibitorscreening assay measures the peroxidase component of cyclooxygenases.The peroxidase activity is assayed colorimetrically by monitoring theappearance of oxidized N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD).This inhibitor screening assay includes both COX-1 and COX-2 enzymes inorder to screen isozyme-specific inhibitors. The Colormetric COX (ovine)Inhibitor screening assay (#760111, Cayman Chemical) can be used toanalyze the effects of each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification on the activity of purifiedcyclooxygenase enzyme (COX-1 or COX-2). According to manufacturerinstructions, purified enzyme, heme and test extracts can be mixed inassay buffer and incubated with shaking for 15 min at room temperature.Following incubation, arachidonic acid and colorimetric substrate can beadded to initiate the reaction. Color progression can be evaluated bycolorimetric plate reading at 590 nm. The percent inhibition of COX-1 orCOX-2 activity can be calculated compared to non-treated controls todetermine the ability of test extracts to inhibit the activity ofpurified enzyme.

Lipoxygenase (LO) Assay: An in vitro lipoxygenase (LO) inhibition assay.LOs are non-heme iron-containing dioxygenases that catalyze the additionof molecular oxygen to fatty acids. Linoleate and arachidonate are themain substrates for LOs in plants and animals. Arachidonic acid may thenbe converted to hydroxyeicosotrienenoic (HETE) acid derivatives, thatare subsequently converted to leukotrienes, potent inflammatorymediators. This assay provides an accurate and convenient method forscreening lipoxygenase inhibitors by measuring the hydroperoxidesgenerated from the incubation of a lipoxygenase (5-, 12-, or 15-LO) witharachidonic acid. The Colorimetric LO Inhibitor screening kit (#760700,Cayman Chemical) can be used to determine the ability of each of theactive ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification toinhibit enzyme activity. Purified 15-lipoxygenase and test ingredientscan be mixed in assay buffer and incubated with shaking for 10 min atroom temperature. Following incubation, arachidonic acid can be added toinitiate the reaction and the mixtures can be incubated for anadditional 10 min at room temperature. Colorimetric substrate can beadded to terminate catalysis and color progression can be evaluated byfluorescence plate reading at 490 nm. The percent inhibition oflipoxyganse activity can be calculated compared to non-treated controlsto determine the ability of each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification to inhibit the activity of purifiedenzyme.

B16 Pigmentation Assay: Melanogenesis is the process by whichmelanocytes produce melanin, a naturally produced pigment that impartscolor to skin, hair, and eyes. Inhibiting melanogenesis is beneficial toprevent skin darkening and lighten dark spots associated with aging.

This bioassay utilizes B16-F1 melanocytes (ATCC), an immortalized mousemelanoma cell line, to analyze the effect of compounds on melanogenesis.The endpoint of this assay is a spectrophotometric measurement ofmelanin production and cellular viability. B16-F1 melanocytes can becultivated in standard DMEM growth medium with 10% fetal bovine serum(Mediatech) at 37° C. in 10% CO₂ and then treated with any one of theactive ingredients, combination of ingredients, or compositions havingsaid combinations disclosed in the specification for 6 days. Followingincubation, melanin secretion is measured by absorbance at 405 nm andcellular viability was quantified.

Elastin Stimulation Assay: Elastin is a connective tissue protein thathelps skin resume shape after stretching or contracting. Elastin is alsoan important load-bearing protein used in places where mechanical energyis required to be stored. Elastin is made by linking many solubletropoelastin protein molecules, in a reaction catalyzed by lysyloxidase. Elastin secretion and elastin fibers can be monitored incultured human fibroblasts by staining of cultured human fibroblastsusing immunofluorescent antibodies directed against elastin.

Laminin and Fibronectin Stimulation Assay: Laminin and fibronectin aremajor proteins in the dermal-epidermal junction (DEJ) (also referred toas the basement membrane). The DEJ is located between the dermis and theepidermis and interlocks forming fingerlike projections called reteridges. The cells of the epidermis receive their nutrients from theblood vessels in the dermis. The rete ridges increase the surface areaof the epidermis that is exposed to these blood vessels and the needednutrients. The DEJ provides adhesion of the two tissue compartments andgoverns the structural integrity of the skin. Laminin and fibronectinare two structural glycoproteins located in the DEJ. Considered the gluethat holds the cells together, laminin and fibronectin are secreted bydermal fibroblasts to help facilitate intra- and inter-cellular adhesionof the epidermal cells to the DEJ. Laminin and fibronectin secretion canbe monitored by quantifying laminin and fibronectin in cell supernatantsof cultured human fibroblasts treated for 3 days with culture mediumwith or without the test ingredient(s). Following incubation, lamininand fibronectin content can be measured using immunofluorescentantibodies directed against each protein in an enzyme linkedimmuno-sorbant assay (ELISA). Measurements are normalized for cellularmetabolic activity, as determined by bioconversion of3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS).

Tumor Necrosis Factor Alpha (TNF-α) Assay: The prototype ligand of theTNF superfamily, TNF-α, is a pleiotropic cytokine that plays a centralrole in inflammation. Increase in its expression is associated with anup regulation in pro-inflammatory activity. This bioassay can be used toanalyze the effect of any one of the active ingredients, combination ofingredients, or compositions having said combinations disclosed in thespecification on the production of TNF-α by human epidermalkeratinocytes. The endpoint of this assay can be a spectrophotometricmeasurement that reflects the presence of TNF-α and cellular viability.The assay employs the quantitative sandwich enzyme immunoassay techniquewhereby a monoclonal antibody specific for TNF-α has been pre-coatedonto a microplate. Standards and samples can be pipetted into the wellsand any TNF-α present is bound by the immobilized antibody. Afterwashing away any unbound substances, an enzyme-linked polyclonalantibody specific for TNF-α can be added to the wells. Following a washto remove any unbound antibody-enzyme reagent, a substrate solution canbe added to the wells and color developed in proportion to the amount ofTNF-α bound in the initial step using a microplate reader for detectionat 450 nm. The color development can be stopped and the intensity of thecolor can be measured. Subconfluent normal human adult keratinocytes(Cascade Biologics) cultivated in EPILIFE™ standard growth medium(Cascade Biologics) at 37° C. in 5% CO₂, can be treated with phorbol12-myristate 13-acetate (PMA , 10 ng/ml, Sigma Chemical, #P1585-1MG) andany one of the active ingredients, combination of ingredients, orcompositions having said combinations disclosed in the specification for6 hours. PMA has been shown to cause a dramatic increase in TNF-αsecretion which peaks at 6 hours after treatment. Following incubation,cell culture medium can be collected and the amount of TNF-α secretionquantified using a sandwich enzyme linked immuno-sorbant assay (ELISA)from R&D Systems (#DTA00C).

Mushroom tyrosinase activity assay: In mammalian cells, tyrosinasecatalyzes two steps in the multi-step biosynthesis of melanin pigmentsfrom tyrosine (and from the polymerization of dopachrome). Tyrosinase islocalized in melanocytes and produces melanin (aromatic quinonecompounds) that imparts color to skin, hair, and eyes. Purified mushroomtyrosinase (Sigma) can be incubated with its substrate L-Dopa (Fisher)in the presence or absence of each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification. Pigment formation can be evaluated bycolorimetric plate reading at 490 nm. The percent inhibition of mushroomtyrosinase activity can be calculated compared to non-treated controlsto determine the ability of test ingredients or combinations thereof toinhibit the activity of purified enzyme. Test extract inhibition can becompared with that of kojic acid (Sigma).

Elastase Assay: ENZCHEK® Elastase Assay (Kit# E-12056) from MolecularProbes (Eugene, Oreg. USA) can be used as an in vitro enzyme inhibitionassay for measuring inhibition of elastase activity for each of theactive ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification.The EnzChek kit contains soluble bovine neck ligament elastin that canbe labeled with dye such that the conjugate's fluorescence can bequenched. The non-fluorescent substrate can be digested by elastase orother proteases to yield highly fluorescent fragments. The resultingincrease in fluorescence can be monitored with a fluorescence microplatereader. Digestion products from the elastin substrate have absorptionmaxima at ˜505 nm and fluorescence emission maxima at ˜515 nm. Thepeptide, N-methoxysuccinyl-Ala-Ala-Pro-Val- chloromethyl ketone, can beused as a selective, collective inhibitor of elastase when utilizing theEnzChek Elastase Assay Kit for screening for elastase inhibitors.

Oil Control Assay: An assay to measure reduction of sebum secretion fromsebaceous glands and/or reduction of sebum production from sebaceousglands can be assayed by using standard techniques known to those havingordinary skill in the art. In one instance, the forehead can be used.Each of the active ingredients, any one of the combination ofingredients, or compositions having said combinations disclosed in thespecification can be applied to one portion of the forehead once ortwice daily for a set period of days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, or more days), while another portion of the foreheadis not treated with the composition. After the set period of daysexpires, then sebum secretion can be assayed by application of fineblotting paper to the treated and untreated forehead skin. This is doneby first removing any sebum from the treated and untreated areas withmoist and dry cloths. Blotting paper can then be applied to the treatedand untreated areas of the forehead, and an elastic band can be placedaround the forehead to gently press the blotting paper onto the skin.After 2 hours the blotting papers can be removed, allowed to dry andthen transilluminated. Darker blotting paper correlates with more sebumsecretion (or lighter blotting paper correlates with reduced sebumsecretion.

Erythema Assay: An assay to measure the reduction of skin redness can beevaluated using a Minolta Chromometer. Skin erythema may be induced byapplying a 0.2% solution of sodium dodecyl sulfate on the forearm of asubject. The area is protected by an occlusive patch for 24 hrs. After24 hrs, the patch is removed and the irritation-induced redness can beassessed using the a* values of the Minolta Chroma Meter. The a* valuemeasures changes in skin color in the red region. Immediately afterreading, the area is treated with the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification. Repeat measurements can be taken atregular intervals to determine the formula's ability to reduce rednessand irritation.

Skin Moisture/Hydration Assay: Skin moisture/hydration benefits can bemeasured by using impedance measurements with the Nova Dermal PhaseMeter. The impedance meter measures changes in skin moisture content.The outer layer of the skin has distinct electrical properties. Whenskin is dry it conducts electricity very poorly. As it becomes morehydrated increasing conductivity results. Consequently, changes in skinimpedance (related to conductivity) can be used to assess changes inskin hydration. The unit can be calibrated according to instrumentinstructions for each testing day. A notation of temperature andrelative humidity can also be made. Subjects can be evaluated asfollows: prior to measurement they can equilibrate in a room withdefined humidity (e.g., 30-50%) and temperature (e.g., 68-72° C.). Threeseparate impedance readings can be taken on each side of the face,recorded, and averaged. The T5 setting can be used on the impedancemeter which averages the impedance values of every five secondsapplication to the face. Changes can be reported with statisticalvariance and significance. Each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification can be assayed according to this process.

Skin Clarity and Reduction in Freckles and Age Spots Assay: Skin clarityand the reduction in freckles and age spots can be evaluated using aMinolta Chromometer. Changes in skin color can be assessed to determineirritation potential due to product treatment using the a* values of theMinolta Chroma Meter. The a* value measures changes in skin color in thered region. This is used to determine whether each of the activeingredients, any one of the combination of ingredients, or compositionshaving said combinations disclosed in the specification is inducingirritation. The measurements can be made on each side of the face andaveraged, as left and right facial values. Skin clarity can also bemeasured using the Minolta Meter. The measurement is a combination ofthe a*, b, and L values of the Minolta Meter and is related to skinbrightness, and correlates well with skin smoothness and hydration. Skinreading is taken as above. In one non-limiting aspect, skin clarity canbe described as L/C where C is chroma and is defined as (a²+b2)^(1/2).

Skin Dryness, Surface Fine Lines, Skin Smoothness, and Skin Tone Assay:Skin dryness, surface fine lines, skin smoothness, and skin tone can beevaluated with clinical grading techniques. For example, clinicalgrading of skin dryness can be determined by a five point standardKligman Scale: (0) skin is soft and moist; (1) skin appears normal withno visible dryness; (2) skin feels slightly dry to the touch with novisible flaking; (3) skin feels dry, tough, and has a whitish appearancewith some scaling; and (4) skin feels very dry, rough, and has a whitishappearance with scaling. Evaluations can be made independently by twoclinicians and averaged.

Clinical Grading of Skin Tone Assay: Clinical grading of skin tone canbe performed via a ten point analog numerical scale: (10) even skin ofuniform, pinkish brown color. No dark, erythremic, or scaly patches uponexamination with a hand held magnifying lens. Microtexture of the skinvery uniform upon touch; (7) even skin tone observed withoutmagnification. No scaly areas, but slight discolorations either due topigmentation or erythema. No discolorations more than 1 cm in diameter;(4) both skin discoloration and uneven texture easily noticeable. Slightscaliness. Skin rough to the touch in some areas; and (1) uneven skincoloration and texture. Numerous areas of scaliness and discoloration,either hypopigmented, erythremic or dark spots. Large areas of unevencolor more than 1 cm in diameter. Evaluations were made independently bytwo clinicians and averaged.

Clinical Grading of Skin Smoothness Assay: Clinical grading of skinsmoothness can be analyzed via a ten point analog numerical scale: (10)smooth, skin is moist and glistening, no resistance upon dragging fingeracross surface; (7) somewhat smooth, slight resistance; (4) rough,visibly altered, friction upon rubbing; and (1) rough, flaky, unevensurface. Evaluations were made independently by two clinicians andaveraged.

Skin Smoothness and Wrinkle Reduction Assay With Methods Disclosed inPackman et al. (1978): Skin smoothness and wrinkle reduction can also beassessed visually by using the methods disclosed in Packman et al.(1978). For example, at each subject visit, the depth, shallowness andthe total number of superficial facial lines (SFLs) of each subject canbe carefully scored and recorded. A numerical score was obtained bymultiplying a number factor times a depth/width/length factor. Scoresare obtained for the eye area and mouth area (left and right sides) andadded together as the total wrinkle score.

Skin Firmness Assay with a Hargens Ballistometer: Skin firmness can bemeasured using a Hargens ballistometer, a device that evaluates theelasticity and firmness of the skin by dropping a small body onto theskin and recording its first two rebound peaks. The ballistometry is asmall lightweight probe with a relatively blunt tip (4 square mm-contactarea) was used. The probe penetrates slightly into the skin and resultsin measurements that are dependent upon the properties of the outerlayers of the skin, including the stratum corneum and outer epidermisand some of the dermal layers.

Skin Softness/Suppleness Assay with a Gas Bearing Electrodynamometer:Skin softness/suppleness can be evaluated using the Gas BearingElectrodynamometer, an instrument that measures the stress/strainproperties of the skin. The viscoelastic properties of skin correlatewith skin moisturization. Measurements can be obtained on thepredetermined site on the cheek area by attaching the probe to the skinsurface with double-stick tape. A force of approximately 3.5 gm can beapplied parallel to the skin surface and the skin displacement isaccurately measured. Skin suppleness can then be calculated and isexpressed as DSR (Dynamic Spring Rate in gm/mm).

Appearance of Lines and Wrinkles Assay with Replicas: The appearance oflines and wrinkles on the skin can be evaluated using replicas, which isthe impression of the skin's surface. Silicone rubber like material canbe used. The replica can be analyzed by image analysis. Changes in thevisibility of lines and wrinkles can be objectively quantified via thetaking of silicon replicas form the subjects' face and analyzing thereplicas image using a computer image analysis system. Replicas can betaken from the eye area and the neck area, and photographed with adigital camera using a low angle incidence lighting. The digital imagescan be analyzed with an image processing program and are of the replicascovered by wrinkles or fine lines was determined.

Surface Contour of the Skin Assay with a Profilometer/Stylus Method: Thesurface contour of the skin can be measured by using theprofilometer/Stylus method. This includes either shining a light ordragging a stylus across the replica surface. The vertical displacementof the stylus can be fed into a computer via a distance transducer, andafter scanning a fixed length of replica a cross-sectional analysis ofskin profile can be generated as a two-dimensional curve. This scan canbe repeated any number of times along a fix axis to generate a simulated3-D picture of the skin. Ten random sections of the replicas using thestylus technique can be obtained and combined to generate averagevalues. The values of interest include Ra which is the arithmetic meanof all roughness (height) values computed by integrating the profileheight relative to the mean profile height. Rt which is the maximumvertical distance between the highest peak and lowest trough, and Rzwhich is the mean peak amplitude minus the mean peak height. Values aregiven as a calibrated value in mm. Equipment should be standardizedprior to each use by scanning metal standards of know values. Ra Valuecan be computed by the following equation: R_(a)=Standardize roughness;l_(m)=the traverse (scan) length; and y=the absolute value of thelocation of the profile relative to the mean profile height (x-axis).

MELANODERM™ Assay: In other non-limiting aspects, the efficacy of eachof the active ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification canbe evaluated by using a skin analog, such as, for example, MELANODERM™.Melanocytes, one of the cells in the skin analog, stain positively whenexposed to L-dihydroxyphenyl alanine (L-DOPA), a precursor of melanin.The skin analog, MELANODERM™, can be treated with a variety of basescontaining each of the active ingredients, any one of the combination ofingredients, or compositions having said combinations disclosed in thespecification or with the base alone as a control. Alternatively, anuntreated sample of the skin analog can be used as a control.

Production of Filaggrin: Changes in the production of filaggrin inkeratinocytes due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Filaggrin is theprecursor to Natural Moisturizing Factor (NMF) in the skin. IncreasedNNW increases the moisture content of the skin. Filaggrin production intreated and non-treated keratinocytes can be determined using a bioassaythat analyzes filaggrin concentration in keratinocyte cell lysates. Anon-limiting example of a bioassay that can be used to quantifyfilaggrin production is the PROTEINSIMPLE® SIMON™ western blottingprotocol. For each sample, normal human epidermal keratinocytes (NHEK)are grown in EPI-200—Mattek EPILIFE™ growth media with calcium from LifeTechnologies (M-EP-500-CA). NHEK are incubated in growth mediumovernight at 37° C. in 5% CO₂ prior to treatment. NHEK are thenincubated in growth medium with 1% test compound/extract or nocompound/extract (negative control) for 24 to 36 hours. The NHEK canthen be washed, collected, and stored on ice or colder until lysed onice using a lysis buffer and sonication. The protein concentrations ofthe samples can be determined and used to normalize the samples. Thelysates can be stored at —80° C. until use in the quantification assay.

The PROTEINSIMPLE® SIMON™ western blotting bioassay assay employs aquantitative western blotting immunoassay technique using an antibodyspecific for filaggrin to quantitatively detect filaggrin in the testsamples. Cell samples are lysed and normalized for proteinconcentration. Normalized samples and molecular weight standards canthen be loaded and ran on a denatured protein separation gel usingcapillary electrophoresis. The proteins in the gel are immobilized andimmunoprobed using a primary antibody specific for filaggrin. Theimmobilized proteins can then be immunoprobed with an enzyme-linkeddetection antibody that binds the primary antibody. A chemiluminescentsubstrate solution can then be added to the immobilized proteins toallow chemiluminescent development in proportion to the amount offilaggrin bound in the immobilization. The chemiluminescent developmentis stopped at a specific time and the intensity of the chemiluminescentsignal can be measured and compared to positive and negative controls.

Production of Occludin: Changes in the production of occludin inkeratinocytes due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Occludin is a proteincritical to the formulation of tight junctions and the skin's moisturebarrier function. A non-limiting example of how occludin production intreated and non-treated keratinocytes can be determined is by the use ofa bioassay that analyzes occludin concentration in keratinocyte celllysates. The bioassay can be performed using PROTEINSIMPLE® SIMON™western blotting protocol. For the samples, adult human epidermalkeratinocytes (HEKa) from Life Technologies (C-005-5C) can be grown at37° C. and 5% CO₂ for 24 hours in EPILIFE™ growth media with calciumfrom Life Technologies (M-EP-500-CA) supplemented with KeratinocyteGrowth Supplement (HKGS) from Life Technologies (S-101-5). HEKa are thenincubated in growth medium with test compound/extract, nocompound/extract for negative control, or with 1 mM CaCl₂ for positivecontrol for 24 to 48 hours. The HEKa are then washed, collected, andstored on ice or colder until lysed on ice using a lysis buffer andsonication. The protein concentrations of the samples can be determinedand used to normalize the samples. The lysates are stored at —80° C.until use in the bioassay.

The PROTEINSIMPLE® SIMON™ western blotting bioassay assay employs aquantitative western blotting immunoassay technique using an antibodyspecific for occludin to quantitatively detect occludin in the testsamples. Cell samples are lysed and normalized for proteinconcentration. Normalized samples and molecular weight standards arethen loaded and ran on a denatured protein separation gel usingcapillary electrophoresis. The proteins in the gel are then immobilizedand immunoprobed using a primary antibody specific for occludin. Theimmobilized proteins are immunoprobed with an enzyme-linked detectionantibody that binds the primary antibody. A chemiluminescent substratesolution is then added to the immobilized proteins to allowchemiluminescent development in proportion to the amount of occludinbound in the immobilization. The chemiluminescent development can bestopped at a specific time and the intensity of the chemiluminescentsignal can be measured and compared to positive and negative controls.

Keratinocyte Monolayer Permeability: Changes in the permeability of akeratinocyte monolayer due to each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Keratinocyte monolayerpermeability is a measure of skin barrier integrity. Keratinocytemonolayer permeability in treated and non-treated keratinocytes can bedetermined using, as a non-limiting example, the In Vitro VascularPermeability assay by Millipore (ECM642). This assay analyzesendothelial cell adsorption, transport, and permeability. Briefly, adulthuman epidermal keratinocytes from Life Technologies (C-005-5C) can beseeded onto a porous collagen-coated membrane within a collection well.The keratinocytes are then incubated for 24 hours at 37° C. and 5% CO₂in EPILIFE™ growth media with calcium from Life Technologies(M-EP-500-CA) supplemented with Keratinocyte Growth Supplement (HKGS)from Life Technologies (S-101-5). This incubation time allows the cellsto form a monolayer and occlude the membrane pores. The media is thenreplaced with fresh media with (test sample) or without (non-treatedcontrol) test compounds/extracts and the keratinocytes are incubated foran additional 48 hours at 37° C. and 5% CO₂. To determine permeabilityof the keratinocyte monolayer after incubation with/without the testcompound/extract, the media is replaced with fresh media containing ahigh molecular weight Fluorescein isothiocyanate (FITC)-Dextran and thekeratinocytes are incubated for 4 hours at 37° C. and 5% CO₂. During the4 hours incubation, FITC can pass through the keratinocytes monolayerand porous membrane into the collection well at a rate proportional tothe monolayer's permeability. After the 4 hour incubation, cellviability and the content of FITC in the collection wells can bedetermined. For the FITC content, the media in the collection well iscollected and fluorescence of the media determined at 480 nm (Em) whenexcited at 520 nm. Percent permeability and percent change in comparisonto the non-treated controls can be determined by the followingequations: Percent Permeability=((Mean Ex/Em of test sample)/Mean Ex/Emuntreated control)*100; Percent Change=Percent Permeability of testsample−Percent Permeability of untreated control.

Production of Hyaluronic Acid: Changes in the production of hyaluronicacid in human dermal fibroblasts due to each of the active ingredients,any one of the combination of ingredients, or compositions having saidcombinations disclosed in the specification can be measured. HA is apolysaccharide involved in stabilization of the structure of the matrixand is involved in providing turgor pressure to tissue and cells. As onenon-limiting example, HA production in treated and non-treated adulthuman dermal fibroblasts (HDFa) cells can be determined using theHyaluronan DuoSet ELISA kit from R&D Systems (DY3614). In this assay,for production of samples, subconfluent HDFa cells from CascadeBiologics (C-13-5C) are incubated at 37° C. and 10% CO₂ in starvationmedium (0.15% fetal bovine serum and 1% Penicillin Streptomycin solutionin Dulbecco's Modified Eagle Medium) for 72 hours prior to treatment.The cells are then incubated with fresh starvation medium with eithertest compound, positive control (phorbol 12-myristate 13-acetate fromSigma-Aldrich (P1585) and platelet derived growth factor fromSigma-Aldrich (P3201)), or no additive for 24 hours. Media is thencollected and frozen at −80° C. until use in the ELISA assay.

Briefly, the ELISA assay employs a quantitative sandwich enzymeimmunoassay technique whereby a capture antibody specific for HA can bepre-coated onto a microplate. Standards and media from treated anduntreated cells are pipetted into the microplate wells to enable any HApresent to be bound by the immobilized antibody. After washing away anyunbound substances, an enzyme-linked detection antibody specific for HAis added to the wells. Following a wash to remove any unboundantibody-enzyme reagent, a substrate solution is added to the wells toallow color development in proportion to the amount of HA bound in theinitial step. The color development is stopped at a specific time andthe intensity of the color at 450 nm can be measured using a microplatereader.

Inhibition of Hyaluronidase Activity: Changes in the activity ofhyaluronidase due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Hyaluronidase is anenzyme that degrades HA. HA is a polysaccharide involved instabilization of the structure of the matrix and is involved inproviding turgor pressure to tissue and cells. As one non-limitingexample, hyaluronidase activity can be determined using an in vitroprotocol modified from Sigma-Aldrich protocol # EC 3.2.1.35. Briefly,hyaluronidase type 1-S from Sigma-Aldrich (H3506) is added to microplatereaction wells containing test compound or controls. Tannic acid can beused as a positive control inhibitor, no test compound can be added forthe control enzyme, and wells with test compound or positive control butwithout hyaluronidase can be used as a background negative control. Thewells are incubated at 37° C. for 10 minutes before addition ofsubstrate (HA). Substrate is added and the reactions incubated at 37° C.for 45 minutes. A portion of each reaction solution is then transferredto and gently mixed in a solution of sodium acetate and acetic acid pH3.75 to stop that portion of the reaction (stopped wells). The stoppedwells and the reaction wells should both contain the same volume ofsolution after addition of the portion of the reaction solution to thestopped wells. Both the reaction wells and the stopped wells areincubated for 10 minutes at room temperature. Absorbance at 600 nm isthen measured for both the reaction wells and the stopped wells.Inhibition can be calculated using the following formulas: Inhibitor (orcontrol) activity=(Inhibitor stopped wells absorbance at 600nm−inhibitor reaction wells absorbance at 600 nm); Initialactivity=control enzyme absorbance at 600 nm; Percent Inhibition=[(Initial activity/Inhibitor Activity)*100]−100.

Peroxisome Proliferator-Activated Receptor Gamma (PPAR-γ) Activity:Changes in the activity of PPAR-γ due to each of the active ingredients,any one of the combination of ingredients, or compositions having saidcombinations disclosed in the specification can be measured. PPAR-γ is areceptor critical for the production of sebum. As one non-limitingexample, the activity of PPAR-γ can be determined using a bioassay thatanalyzes the ability of a test compound or composition to inhibitbinding of a ligand. Briefly, fluorescent small-molecule pan-PPARligand, FLUORMONE™ Pan-PPAR Green, available from Life Technologies(PV4894), can be used to determine if test compounds or compositions areable to inhibit binding of the ligand to PPAR-γ. The samples wellsinclude PPAR-γ and fluorescent ligand and either: test compound orcomposition (test); a reference inhibitor, rosiglitazone (positivecontrol); or no test compound (negative control). The wells areincubated for a set period of time to allow the ligand opportunity tobind the PPAR-γ. The fluorescence polarization of each sample well canthen be measured and compared to the negative control well to determinethe percentage of inhibition by the test compound or composition.

Cytokine array: Human epidermal keratinocytes are cultured to 70-80%confluency. The media in the plate is aspirated and 0.025% trypsin/EDTAis added. When the cells became rounded, the culture dish is gentlytapped to release the cells. The trypsin/EDTA containing cells areremoved from the culture dish and neutralized. Cells are centrifuged for5 min. at 180×g to form a pellet of cells. The supernatant is aspirated.The resulting pellet is resuspended in EPILIFE™ media (CascadeBiologics). The cells are seeded in 6-well plates at approximately10-20% confluency. After the cells became approximately 80% confluent,the media is aspirated and 1.0 ml of EPILIFE™, along with phorbol13-Myristate 12-acetate (“PMA”) (a known inducer of inflammation) andthe test composition dilutions are added to two replicate wells (i.e.,1.0% (100 μl of 100× stock) and 0.1% (10 μl of 100× stock) testcompositions are diluted into a final volume of 1 ml EPILIFE™ GrowthMedium). The media is gently swirled to ensure adequate mixing. Inaddition, 1.0 ml of EPILIFE™ is added to the control wells, with andwithout additional PMA. The plates are then incubated at 37±1° C. and5.0±1% CO₂ for approximately 5 hours after dosing. Following this 5-hourincubation, all media is collected in conical tubes and frozen at −70°C.

For analysis, a 16-pad hybridization chamber is attached to 16-pad FASTslides arrayed in triplicate with 16 anti-cytokine antibodies plusexperimental controls (Whatman BioSciences), and the slides are placedinto a FASTFrame (4 slides per frame) for processing. Arrays are blockedfor 15 min. at room temp. using 70 ml S&S Protein Array Blocking buffer(Whatman Schleicher and Scheull). Blocking buffer is removed and 70 mlof each supernatant sample is added to each array. Arrays are incubatedfor 3 hours at room temp. with gentle agitation. Arrays are washed 3times with TBS-T. Arrays are treated with 70 ml of an antibody cocktail,containing one biotinylated antibody corresponding to each of thearrayed capture antibodies. Arrays are incubated for 1 hour at roomtemp. with gentle agitation. Arrays are washed 3 times with TBS-T.Arrays are incubated with 70 ml of a solution containingstreptavidin-Cy5 conjugate for 1 hour at room temp. with gentleagitation. Arrays are washed 3 times with TBS-T, quickly rinsed inde-ionized water, and dried.

Slides can be imaged in a Perkin-Elmer ScanArray 4000 confocalfluorescent imaging system. Array images can be saved and analyzed usingImaging Research ArrayVision software. Briefly, spot intensities aredetermined by subtracting background signal. Spot replicates from eachsample condition can be averaged and then compared to the appropriatecontrols.

Endothelial Tube Formation: Endothelial tube formation is involved inangiogenesis and micro-vessel capillary formation. Capillary formationand angiogenesis may contribute to redness and rosacea of the skin. Theability for endothelial cells to form tubes in the presence or absenceof test extracts and compounds may be determined using a capillarytubule disruption assay with pre-formed primary human umbilical veinendothelial cells (HUVEC) in a cell culture system.

Briefly, HUVECs are cultured in vitro on Extracellular Matrix, whichstimulates the attachment and tubular morphogenesis of endothelial cellsto form capillary-like lumen structures. These in vitro formed capillarytubules are similar to human blood vessel capillaries in many aspects.The capillary tube assay is based on this phenomenon and is used forevaluation of potential vasculature targeting agents.

HUVEC cultures are grown in a 5% CO₂ 37° C. cell incubator. The fullgrowth medium for HUVECs is Endothelial Cell Basal Medium (EBM)supplemented with 2% fetal bovine serum (FBS), 12 μg /ml bovine brainextract, 1 μg/ml hydrocortisone, and 1 μg/ml GA-1000(gentamicin-amphothericin). HUVEC cultures between passage 3 and 8 maybe used for all assay experiments.

HUVECs are pre-labeled with fluorescent agent Calcein AM and seeded inExtracellular Matrix coated 96-well culture plate with their full growthmedium. After about four hours of the morphogenesis process, theendothelial capillary tubes should be formed. Then, test agent indesigned doses in 50 μl volume is applied into the formed capillarytubule cultures as treatment conditions. The no-treatment controls canbe added with vehicle of test agents. Sutent, a FDA approvedanti-angiogenic drug one concentration can be included as assayperformance control. After about six hours of treatment, the endothelialtubule morphology in each well is examined by microscopy, imaged, andthe capillary disrupting activities under treatment conditions can bequantitatively analyzed. Each test conditions can be conducted induplicate wells, including controls.

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

1. A method for reducing muscle contraction of a facial muscle in aperson and/or reducing the appearance of a fine line or wrinkle on aperson's skin, the method comprising topically applying to facial skin acomposition comprising: (a) an effective amount of Rosmarinusofficinalis leaf extract; (b) an effective amount of Lavendula stoechasextract; and (c) an effective amount of Acmella oleracea extract,wherein topical application of the composition to the facial skinreduces muscle contraction of the facial muscle and/or reduces theappearance of the fine line or wrinkle.
 2. The method of claim 1,wherein the composition is applied to a fine line or wrinkle, andwherein the appearance of the fine line or wrinkle is reduced aftertopical application and/or muscle contraction of a muscle that causesthe appearance of the fine line or wrinkle is reduced after topicalapplication.
 3. The method of claim 2, wherein the fine line or wrinkleis a rhytide, and wherein the reduction in muscle contraction of thefacial muscle reduces the appearance of the rhytide.
 4. The method ofclaim 1, wherein the facial muscle is a glabellar complex muscle, anorbicularis oculi muscle, a depressor muscle, or a frontalis muscle, orany combination thereof.
 5. The method of claim 1, wherein each of theextracts is individually an aqueous extract, an alcohol extract, apolyol extract, or a combination thereof.
 6. The method of claim 1,wherein: the Rosmarinus officinalis leaf extract is obtained with afluid extraction solvent mixture comprising betaine, lactic acid, andwater; the Lavendula stoechas extract is a Lavendula stoechasflower/leaf/stem extract obtained with a supercritical carbon dioxide(CO₂) extraction solvent; and the Acmella oleracea extract is obtainedwith a fluid extraction solvent mixture comprising water, ethanol, and1,3 propanediol.
 7. The method of claim 1, wherein the compositioncomprises: 0.00001 to 10% w/w of Rosmarinus officinalis leaf extract;0.00001 to 10% w/w of Lavendula stoechas extract; and, 0.00001 to 10%w/w of Acmella oleracea extract.
 8. The method of claim 1, wherein: theRosmarinus officinalis leaf extract reduces an influx of calcium in thefacial muscle and reduces the occurrence of an action potential in thefacial muscle; the Lavendula stoechas extract reduces an influx ofcalcium in the facial muscle and reduces the occurrence of an actionpotential in the facial muscle; and the Acmella oleracea extract reducesan influx of calcium in the facial muscle and reduces the occurrence ofan action potential in the facial muscle.
 9. The method of claim 1,wherein the Acmella oleracea extract stimulates collagen production andlaminin production in the skin.
 10. The method of claim 1, wherein theRosmarinus officinalis leaf extract reduces matrix metalloproteinases 1,3, or 9 production in the skin.
 11. The method of claim 1, wherein thefacial skin is forehead skin, cheek skin, chin skin, and/or orbital areaskin.
 12. The method of claim 1, wherein the composition is an emulsion.13. The method of claim 12, wherein the emulsion is an oil-in-wateremulsion.
 14. The method of claim 1, wherein the composition is a gel.15. A topical skin composition comprising: (a) an effective amount ofRosmarinus officinalis leaf extract; (b) an effective amount ofLavendula stoechas extract; and (c) an effective amount of Acmellaoleracea extract, wherein the topical skin composition is capable ofreducing the appearance of a fine line or wrinkle in a person's skinand/or is capable of reducing muscle contraction of a facial muscle in aperson.
 16. The topical skin composition of claim 15, wherein each ofthe extracts is individually an aqueous extract, an alcohol extract, apolyol extract, or a combination thereof.
 17. The topical skincomposition of claim 15, wherein: the Rosmarinus officinalis leafextract is obtained with a fluid extraction solvent mixture comprisingbetaine, lactic acid, and water; the Lavendula stoechas extract is aLavendula stoechas flower/leaf/stem extract obtained with asupercritical carbon dioxide (CO₂) extraction solvent; and the Acmellaoleracea extract is obtained with a fluid extraction solvent mixturecomprising water, ethanol, and 1,3 propanediol.
 18. The topical skincomposition of claim 15, wherein the composition comprises: 0.00001 to10% w/w of Rosmarinus officinalis leaf extract; 0.00001 to 10% w/w ofLavendula stoechas extract; and, 0.00001 to 10% w/w of Acmella oleraceaextract.
 19. The topical skin composition of claim 15, wherein: theRosmarinus officinalis leaf extract is capable of reducing an influx ofcalcium in the facial muscle and reduces the occurrence of an actionpotential in the facial muscle and/or the Rosmarinus officinalis leafextract is capable of reducing matrix metalloproteinases 1, 3, or 9production in the skin; the Lavendula stoechas extract is capable ofreducing an influx of calcium in the facial muscle and reduces theoccurrence of an action potential in the facial muscle; and the Acmellaoleracea extract is capable of reducing an influx of calcium in thefacial muscle and reduces the occurrence of an action potential in thefacial muscle, and/or the Acmella oleracea extract is capable ofstimulating collagen production and laminin production in the skin. 20.The topical skin composition of claim 15, wherein the composition is anemulsion, preferably an oil-in-water emulsion, or a gel.